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Christopher Vincent Nicchitta

Professor of Cell Biology
Cell Biology
Duke Box 3709, Durham, NC 27710
436 Nanaline H Duke, Durham, NC 27708

Selected Publications


High-throughput quantitation of protein-RNA UV-crosslinking efficiencies as a predictive tool for high-confidence identification of RNA-binding proteins.

Journal Article RNA · May 16, 2024 UV-crosslinking has proven to be an invaluable tool for the identification of RNA-protein interactomes. The paucity of methods for distinguishing background from bona fide RNA-protein interactions, however, makes attribution of RNA-binding function on UV-c ... Full text Link to item Cite

An emerging role for the endoplasmic reticulum in stress granule biogenesis.

Journal Article Semin Cell Dev Biol · March 15, 2024 Stress granules (SGs), structurally dynamic, optically resolvable, macromolecular assemblies of mRNAs, RNA binding proteins (RBPs), translation factors, ribosomal subunits, as well as other interacting proteins, assemble in response to cell stress conditio ... Full text Link to item Cite

Examining SRP pathway function in mRNA localization to the endoplasmic reticulum.

Journal Article RNA · November 2023 Signal recognition particle (SRP) pathway function in protein translocation across the endoplasmic reticulum (ER) is well established; its role in RNA localization to the ER remains, however, unclear. In current models, mRNAs undergo translation- and SRP-d ... Full text Link to item Cite

Cooperative regulation of coupled oncoprotein synthesis and stability in triple-negative breast cancer by EGFR and CDK12/13.

Journal Article Proc Natl Acad Sci U S A · September 19, 2023 Evidence has long suggested that epidermal growth factor receptor (EGFR) may play a prominent role in triple-negative breast cancer (TNBC) pathogenesis, but clinical trials of EGFR inhibitors have yielded disappointing results. Using a candidate drug scree ... Full text Link to item Cite

Signal-noise metrics for RNA binding protein identification reveal broad spectrum protein-RNA interaction frequencies and dynamics.

Journal Article Nature communications · September 2023 Recent efforts towards the comprehensive identification of RNA-bound proteomes have revealed a large, surprisingly diverse family of candidate RNA-binding proteins (RBPs). Quantitative metrics for characterization and validation of protein-RNA interactions ... Full text Cite

ABL kinases regulate translation in HER2+ cells through Y-box-binding protein 1 to facilitate colonization of the brain.

Journal Article Cell Rep · August 30, 2022 Patients with human epidermal growth factor receptor 2-positive (HER2+/ERBB2) breast cancer often present with brain metastasis. HER2-targeted therapies have not been successful to treat brain metastases in part due to poor blood-brain barrier (BBB) penetr ... Full text Link to item Cite

Recruitment of endoplasmic reticulum-targeted and cytosolic mRNAs into membrane-associated stress granules.

Journal Article RNA · October 2021 Stress granules (SGs) are membraneless organelles composed of mRNAs and RNA binding proteins which undergo assembly in response to stress-induced inactivation of translation initiation. In general, SG recruitment is limited to a subpopulation of a given mR ... Full text Link to item Cite

Quantitative Proteomics Links the LRRC59 Interactome to mRNA Translation on the ER Membrane.

Journal Article Mol Cell Proteomics · November 2020 Protein synthesis on the endoplasmic reticulum (ER) requires the dynamic coordination of numerous cellular components. Together, resident ER membrane proteins, cytoplasmic translation factors, and both integral membrane and cytosolic RNA-binding proteins o ... Full text Link to item Cite

Acceptability and perceived effectiveness of approaches to support biomedical doctoral student wellness: One size doesn⇔t fit all

Journal Article International Journal of Doctoral Studies · January 1, 2020 Aim/Purpose National and international survey studies have begun to identify heightened levels of depression, anxiety, and burnout among doctoral students. Nevertheless, little research has been done to evaluate which interventions may support doctoral stu ... Full text Cite

Applying the Stress Process Model to Stress-Burnout and Stress-Depression Relationships in Biomedical Doctoral Students: A Cross-Sectional Pilot Study.

Journal Article CBE Life Sci Educ · December 2019 Although doctoral students in the biomedical sciences have been recognized as a population at particular risk for mental health problems such as burnout and depression, little research has been conducted to identify candidate targets for intervention. To t ... Full text Link to item Cite

Burnout and Mental Health Problems in Biomedical Doctoral Students.

Journal Article CBE Life Sci Educ · June 2019 Although burnout and mental health problems may adversely impact quality of scientific research, academic productivity, and attrition in biomedical doctoral training programs, very little research has been done on this topic. Recent studies have used brief ... Full text Link to item Cite

Heterogeneous translational landscape of the endoplasmic reticulum revealed by ribosome proximity labeling and transcriptome analysis.

Journal Article J Biol Chem · May 31, 2019 The endoplasmic reticulum (ER) is a nexus for mRNA localization and translation, and recent studies have demonstrated that ER-bound ribosomes also play a transcriptome-wide role in regulating proteome composition. The Sec61 translocon (SEC61) serves as the ... Full text Link to item Cite

Oncoprotein AEG-1 is an endoplasmic reticulum RNA-binding protein whose interactome is enriched in organelle resident protein-encoding mRNAs.

Journal Article RNA · May 2018 Astrocyte elevated gene-1 (AEG-1), an oncogene whose overexpression promotes tumor cell proliferation, angiogenesis, invasion, and enhanced chemoresistance, is thought to function primarily as a scaffolding protein, regulating PI3K/Akt and Wnt/β-catenin si ... Full text Link to item Cite

Dengue Virus Selectively Annexes Endoplasmic Reticulum-Associated Translation Machinery as a Strategy for Co-opting Host Cell Protein Synthesis.

Journal Article J Virol · April 1, 2018 A primary question in dengue virus (DENV) biology is the molecular strategy for recruitment of host cell protein synthesis machinery. Here, we combined cell fractionation, ribosome profiling, and transcriptome sequencing (RNA-seq) to investigate the subcel ... Full text Open Access Link to item Cite

Flavivirus RNA transactions from viral entry to genome replication.

Journal Article Antiviral Res · October 2016 Upon release of the ∼11 kb single-stranded positive polarity dengue virus genomic RNA (gRNA) into the cytoplasm of an infected cell, it serves as the template for translation of the viral polyprotein, which is cleaved into three structural and seven non-st ... Full text Link to item Cite

Complementary Roles of GADD34- and CReP-Containing Eukaryotic Initiation Factor 2α Phosphatases during the Unfolded Protein Response.

Journal Article Mol Cell Biol · July 1, 2016 Phosphorylation of eukaryotic initiation factor 2α (eIF2α) controls transcriptome-wide changes in mRNA translation in stressed cells. While phosphorylated eIF2α (P-eIF2α) attenuates global protein synthesis, mRNAs encoding stress proteins are more efficien ... Full text Open Access Link to item Cite

Methods to Investigate the Regulatory Role of Small RNAs and Ribosomal Occupancy of Plasmodium falciparum.

Journal Article J Vis Exp · December 4, 2015 The genetic variation responsible for the sickle cell allele (HbS) enables erythrocytes to resist infection by the malaria parasite, P. falciparum. The molecular basis of this resistance, which is known to be multifactorial, remains incompletely understood ... Full text Link to item Cite

Simple and inexpensive ribosome profiling analysis of mRNA translation.

Journal Article Methods · December 2015 The development and application of ribosome profiling has markedly advanced our understanding of ribosomes and mRNA translation. The experimental approach, which relies on deep sequencing of ribosome-protected mRNA fragments generated by treatment of polyr ... Full text Open Access Link to item Cite

LOCAL TRANSLATION. Comment on "Principles of ER cotranslational translocation revealed by proximity-specific ribosome profiling".

Journal Article Science · June 12, 2015 Jan et al. (Research Articles, 7 November 2014, p. 716) propose that ribosomes translating secretome messenger RNAs (mRNAs) traffic from the cytosol to the endoplasmic reticulum (ER) upon emergence of the signal peptide and return to the cytosol after term ... Full text Link to item Cite

Diversity and selectivity in mRNA translation on the endoplasmic reticulum.

Journal Article Nat Rev Mol Cell Biol · April 2015 Pioneering electron microscopy studies defined two primary populations of ribosomes in eukaryotic cells: one freely dispersed through the cytoplasm and the other bound to the surface of the endoplasmic reticulum (ER). Subsequent investigations revealed a s ... Full text Link to item Cite

High-throughput quantitation of protein-RNA UV-crosslinking efficiencies as a predictive tool for high-confidence identification of RNA-binding proteins.

Journal Article RNA · May 16, 2024 UV-crosslinking has proven to be an invaluable tool for the identification of RNA-protein interactomes. The paucity of methods for distinguishing background from bona fide RNA-protein interactions, however, makes attribution of RNA-binding function on UV-c ... Full text Link to item Cite

An emerging role for the endoplasmic reticulum in stress granule biogenesis.

Journal Article Semin Cell Dev Biol · March 15, 2024 Stress granules (SGs), structurally dynamic, optically resolvable, macromolecular assemblies of mRNAs, RNA binding proteins (RBPs), translation factors, ribosomal subunits, as well as other interacting proteins, assemble in response to cell stress conditio ... Full text Link to item Cite

Examining SRP pathway function in mRNA localization to the endoplasmic reticulum.

Journal Article RNA · November 2023 Signal recognition particle (SRP) pathway function in protein translocation across the endoplasmic reticulum (ER) is well established; its role in RNA localization to the ER remains, however, unclear. In current models, mRNAs undergo translation- and SRP-d ... Full text Link to item Cite

Cooperative regulation of coupled oncoprotein synthesis and stability in triple-negative breast cancer by EGFR and CDK12/13.

Journal Article Proc Natl Acad Sci U S A · September 19, 2023 Evidence has long suggested that epidermal growth factor receptor (EGFR) may play a prominent role in triple-negative breast cancer (TNBC) pathogenesis, but clinical trials of EGFR inhibitors have yielded disappointing results. Using a candidate drug scree ... Full text Link to item Cite

Signal-noise metrics for RNA binding protein identification reveal broad spectrum protein-RNA interaction frequencies and dynamics.

Journal Article Nature communications · September 2023 Recent efforts towards the comprehensive identification of RNA-bound proteomes have revealed a large, surprisingly diverse family of candidate RNA-binding proteins (RBPs). Quantitative metrics for characterization and validation of protein-RNA interactions ... Full text Cite

ABL kinases regulate translation in HER2+ cells through Y-box-binding protein 1 to facilitate colonization of the brain.

Journal Article Cell Rep · August 30, 2022 Patients with human epidermal growth factor receptor 2-positive (HER2+/ERBB2) breast cancer often present with brain metastasis. HER2-targeted therapies have not been successful to treat brain metastases in part due to poor blood-brain barrier (BBB) penetr ... Full text Link to item Cite

Recruitment of endoplasmic reticulum-targeted and cytosolic mRNAs into membrane-associated stress granules.

Journal Article RNA · October 2021 Stress granules (SGs) are membraneless organelles composed of mRNAs and RNA binding proteins which undergo assembly in response to stress-induced inactivation of translation initiation. In general, SG recruitment is limited to a subpopulation of a given mR ... Full text Link to item Cite

Quantitative Proteomics Links the LRRC59 Interactome to mRNA Translation on the ER Membrane.

Journal Article Mol Cell Proteomics · November 2020 Protein synthesis on the endoplasmic reticulum (ER) requires the dynamic coordination of numerous cellular components. Together, resident ER membrane proteins, cytoplasmic translation factors, and both integral membrane and cytosolic RNA-binding proteins o ... Full text Link to item Cite

Acceptability and perceived effectiveness of approaches to support biomedical doctoral student wellness: One size doesn⇔t fit all

Journal Article International Journal of Doctoral Studies · January 1, 2020 Aim/Purpose National and international survey studies have begun to identify heightened levels of depression, anxiety, and burnout among doctoral students. Nevertheless, little research has been done to evaluate which interventions may support doctoral stu ... Full text Cite

Applying the Stress Process Model to Stress-Burnout and Stress-Depression Relationships in Biomedical Doctoral Students: A Cross-Sectional Pilot Study.

Journal Article CBE Life Sci Educ · December 2019 Although doctoral students in the biomedical sciences have been recognized as a population at particular risk for mental health problems such as burnout and depression, little research has been conducted to identify candidate targets for intervention. To t ... Full text Link to item Cite

Burnout and Mental Health Problems in Biomedical Doctoral Students.

Journal Article CBE Life Sci Educ · June 2019 Although burnout and mental health problems may adversely impact quality of scientific research, academic productivity, and attrition in biomedical doctoral training programs, very little research has been done on this topic. Recent studies have used brief ... Full text Link to item Cite

Heterogeneous translational landscape of the endoplasmic reticulum revealed by ribosome proximity labeling and transcriptome analysis.

Journal Article J Biol Chem · May 31, 2019 The endoplasmic reticulum (ER) is a nexus for mRNA localization and translation, and recent studies have demonstrated that ER-bound ribosomes also play a transcriptome-wide role in regulating proteome composition. The Sec61 translocon (SEC61) serves as the ... Full text Link to item Cite

Oncoprotein AEG-1 is an endoplasmic reticulum RNA-binding protein whose interactome is enriched in organelle resident protein-encoding mRNAs.

Journal Article RNA · May 2018 Astrocyte elevated gene-1 (AEG-1), an oncogene whose overexpression promotes tumor cell proliferation, angiogenesis, invasion, and enhanced chemoresistance, is thought to function primarily as a scaffolding protein, regulating PI3K/Akt and Wnt/β-catenin si ... Full text Link to item Cite

Dengue Virus Selectively Annexes Endoplasmic Reticulum-Associated Translation Machinery as a Strategy for Co-opting Host Cell Protein Synthesis.

Journal Article J Virol · April 1, 2018 A primary question in dengue virus (DENV) biology is the molecular strategy for recruitment of host cell protein synthesis machinery. Here, we combined cell fractionation, ribosome profiling, and transcriptome sequencing (RNA-seq) to investigate the subcel ... Full text Open Access Link to item Cite

Flavivirus RNA transactions from viral entry to genome replication.

Journal Article Antiviral Res · October 2016 Upon release of the ∼11 kb single-stranded positive polarity dengue virus genomic RNA (gRNA) into the cytoplasm of an infected cell, it serves as the template for translation of the viral polyprotein, which is cleaved into three structural and seven non-st ... Full text Link to item Cite

Complementary Roles of GADD34- and CReP-Containing Eukaryotic Initiation Factor 2α Phosphatases during the Unfolded Protein Response.

Journal Article Mol Cell Biol · July 1, 2016 Phosphorylation of eukaryotic initiation factor 2α (eIF2α) controls transcriptome-wide changes in mRNA translation in stressed cells. While phosphorylated eIF2α (P-eIF2α) attenuates global protein synthesis, mRNAs encoding stress proteins are more efficien ... Full text Open Access Link to item Cite

Methods to Investigate the Regulatory Role of Small RNAs and Ribosomal Occupancy of Plasmodium falciparum.

Journal Article J Vis Exp · December 4, 2015 The genetic variation responsible for the sickle cell allele (HbS) enables erythrocytes to resist infection by the malaria parasite, P. falciparum. The molecular basis of this resistance, which is known to be multifactorial, remains incompletely understood ... Full text Link to item Cite

Simple and inexpensive ribosome profiling analysis of mRNA translation.

Journal Article Methods · December 2015 The development and application of ribosome profiling has markedly advanced our understanding of ribosomes and mRNA translation. The experimental approach, which relies on deep sequencing of ribosome-protected mRNA fragments generated by treatment of polyr ... Full text Open Access Link to item Cite

LOCAL TRANSLATION. Comment on "Principles of ER cotranslational translocation revealed by proximity-specific ribosome profiling".

Journal Article Science · June 12, 2015 Jan et al. (Research Articles, 7 November 2014, p. 716) propose that ribosomes translating secretome messenger RNAs (mRNAs) traffic from the cytosol to the endoplasmic reticulum (ER) upon emergence of the signal peptide and return to the cytosol after term ... Full text Link to item Cite

Diversity and selectivity in mRNA translation on the endoplasmic reticulum.

Journal Article Nat Rev Mol Cell Biol · April 2015 Pioneering electron microscopy studies defined two primary populations of ribosomes in eukaryotic cells: one freely dispersed through the cytoplasm and the other bound to the surface of the endoplasmic reticulum (ER). Subsequent investigations revealed a s ... Full text Link to item Cite

De novo translation initiation on membrane-bound ribosomes as a mechanism for localization of cytosolic protein mRNAs to the endoplasmic reticulum.

Journal Article RNA · October 2014 The specialized protein synthesis functions of the cytosol and endoplasmic reticulum compartments are conferred by the signal recognition particle (SRP) pathway, which directs the cotranslational trafficking of signal sequence-encoding mRNAs from the cytos ... Full text Link to item Cite

A novel ribosomopathy caused by dysfunction of RPL10 disrupts neurodevelopment and causes X-linked microcephaly in humans.

Journal Article Genetics · October 2014 Neurodevelopmental defects in humans represent a clinically heterogeneous group of disorders. Here, we report the genetic and functional dissection of a multigenerational pedigree with an X-linked syndromic disorder hallmarked by microcephaly, growth retar ... Full text Link to item Cite

Multifunctional roles for the protein translocation machinery in RNA anchoring to the endoplasmic reticulum.

Journal Article J Biol Chem · September 12, 2014 Signal sequence-encoding mRNAs undergo translation-dependent localization to the endoplasmic reticulum (ER) and at the ER are anchored via translation on Sec61-bound ribosomes. Recent investigations into the composition and membrane association characteris ... Full text Link to item Cite

The unfolded protein response triggers selective mRNA release from the endoplasmic reticulum.

Journal Article Cell · September 11, 2014 The unfolded protein response (UPR) is a stress response program that reprograms cellular translation and gene expression in response to proteotoxic stress in the endoplasmic reticulum (ER). One of the primary means by which the UPR alleviates this stress ... Full text Open Access Link to item Cite

Rare codons regulate KRas oncogenesis.

Journal Article Curr Biol · January 7, 2013 Oncogenic mutations in the small Ras GTPases KRas, HRas, and NRas render the proteins constitutively GTP bound and active, a state that promotes cancer. Ras proteins share ~85% amino acid identity, are activated by and signal through the same proteins, and ... Full text Link to item Cite

Induction of the unfolded protein response drives enhanced metabolism and chemoresistance in glioma cells.

Journal Article PLoS One · 2013 The unfolded protein response (UPR) is an endoplasmic reticulum (ER)-based cytoprotective mechanism acting to prevent pathologies accompanying protein aggregation. It is frequently active in tumors, but relatively unstudied in gliomas. We hypothesized that ... Full text Link to item Cite

Glycoprotein 96 perpetuates the persistent inflammation of rheumatoid arthritis.

Journal Article Arthritis Rheum · November 2012 OBJECTIVE: The mechanisms that contribute to the persistent activation of macrophages in rheumatoid arthritis (RA) are incompletely understood. The aim of this study was to determine the contribution of endogenous gp96 in Toll-like receptor (TLR)-mediated ... Full text Link to item Cite

Translocation of sickle cell erythrocyte microRNAs into Plasmodium falciparum inhibits parasite translation and contributes to malaria resistance.

Journal Article Cell Host Microbe · August 16, 2012 Erythrocytes carrying a variant hemoglobin allele (HbS), which causes sickle cell disease and resists infection by the malaria parasite Plasmodium falciparum. The molecular basis of this resistance, which has long been recognized as multifactorial, remains ... Full text Link to item Cite

Development of a Grp94 inhibitor.

Journal Article J Am Chem Soc · June 13, 2012 Heat shock protein 90 (Hsp90) represents a promising therapeutic target for the treatment of cancer and other diseases. Unfortunately, results from clinical trials have been disappointing as off-target effects and toxicities have been observed. These detri ... Full text Link to item Cite

The enduring enigma of nuclear translation.

Journal Article J Cell Biol · April 2, 2012 Although the physical separation of transcription in the nucleus and translation in the cytoplasm has presided as a fundamental tenet of cell biology for decades, it has not done so without recurring challenges and contentious debate. In this issue, David ... Full text Link to item Cite

Primary role for endoplasmic reticulum-bound ribosomes in cellular translation identified by ribosome profiling.

Journal Article J Biol Chem · February 17, 2012 In eukaryotic cells, the spatial regulation of protein expression is frequently conferred through the coupling of mRNA localization and the local control of translation. mRNA localization to the endoplasmic reticulum (ER) is a prominent example of such reg ... Full text Link to item Cite

Premature translational termination products are rapidly degraded substrates for MHC class I presentation.

Journal Article PLoS One · 2012 Nearly thirty percent of all newly synthesized polypeptides are targeted for rapid proteasome-mediated degradation. These rapidly degraded polypeptides (RDPs) are a source of antigenic substrates for the MHC class I presentation pathway, allowing for immun ... Full text Link to item Cite

Polysome profiling of the malaria parasite Plasmodium falciparum.

Journal Article Mol Biochem Parasitol · September 2011 In the malaria parasite Plasmodium falciparum, global studies of translational regulation have been hampered by the inability to isolate malaria polysomes. We describe here a novel method for polysome profiling in P. falciparum, a powerful approach which a ... Full text Link to item Cite

Hierarchical regulation of mRNA partitioning between the cytoplasm and the endoplasmic reticulum of mammalian cells.

Journal Article Mol Biol Cell · July 15, 2011 The mRNA transcriptome is currently thought to be partitioned between the cytosol and endoplasmic reticulum (ER) compartments by binary selection; mRNAs encoding cytosolic/nucleoplasmic proteins are translated on free ribosomes, and mRNAs encoding topogeni ... Full text Link to item Cite

Out with the old, in with the new? Comparing methods for measuring protein degradation.

Journal Article Cell Biol Int · May 2011 Protein degradation is a critical factor in controlling cellular protein abundance. Here, we compare classical methods for determining protein degradation rates to a novel GFP (green fluorescent protein) fusion protein based method that assesses the intrin ... Full text Link to item Cite

Analyzing mRNA localization to the endoplasmic reticulum via cell fractionation.

Journal Article Methods Mol Biol · 2011 The partitioning of secretory and membrane protein-encoding mRNAs to the endoplasmic reticulum (ER), and their translation on ER-associated ribosomes, governs access to the secretory/exocytic pathways of the cell. As mRNAs encoding secretory and membrane p ... Full text Link to item Cite

Re-examination of CD91 function in GRP94 (glycoprotein 96) surface binding, uptake, and peptide cross-presentation.

Journal Article J Immunol · December 1, 2010 GRP94 (gp96)-peptide complexes can be internalized by APCs and their associated peptides cross-presented to yield activation of CD8(+) T cells. Investigations into the identity (or identities) of GRP94 surface receptors have yielded conflicting results, pa ... Full text Link to item Cite

Gp93, the Drosophila GRP94 ortholog, is required for gut epithelial homeostasis and nutrient assimilation-coupled growth control.

Journal Article Dev Biol · March 15, 2010 GRP94, the endoplasmic reticulum Hsp90, is a metazoan-restricted chaperone essential for early development in mammals, yet dispensable for mammalian cell viability. This dichotomy suggests that GRP94 is required for the functional expression of secretory a ... Full text Link to item Cite

Glioblastoma proto-oncogene SEC61gamma is required for tumor cell survival and response to endoplasmic reticulum stress.

Journal Article Cancer Res · December 1, 2009 Glioblastoma multiforme is the most prevalent type of adult brain tumor and one of the deadliest tumors known to mankind. The genetic understanding of glioblastoma multiforme is, however, limited, and the molecular mechanisms that facilitate glioblastoma m ... Full text Link to item Cite

Efficient cross-priming of antiviral CD8+ T cells by antigen donor cells is GRP94 independent.

Journal Article J Immunol · October 1, 2009 Cross-priming, the activation of naive CD8+ T cells by dendritic cells presenting Ags synthesized by other cells, is believed to play an important role in the generation of antiviral and antitumor responses. The molecular mechanism(s) underlying cross-prim ... Full text Link to item Cite

Heat shock protein 96 is elevated in rheumatoid arthritis and activates macrophages primarily via TLR2 signaling.

Journal Article J Immunol · April 15, 2009 Macrophages are important mediators of chronic inflammation and are prominent in the synovial lining and sublining of patients with rheumatoid arthritis (RA). Recently, we demonstrated increased TLR2 and TLR4 expression and increased response to microbial ... Full text Link to item Cite

Cell biology: How to combat stress.

Journal Article Nature · February 5, 2009 Full text Link to item Cite

Redundancy renders the glycoprotein 96 receptor scavenger receptor A dispensable for cross priming in vivo.

Journal Article Immunology · December 2008 CD8(+) T cells (T(CD8+)) differentiate into effector cells following recognition of specific peptide-major histocompatibility complex (MHC) class I complexes (pMHC-I) on the surface of professional APCs (pAPCs), such as dendritic cells. Antigenic pMHC-I ca ... Full text Link to item Cite

The exception that reinforces the rule: crosspriming by cytosolic peptides that escape degradation.

Journal Article Immunity · June 2008 The nature of crosspriming immunogens for CD8(+) T cell responses is highly controversial. By using a panel of T cell receptor-like antibodies specific for viral peptides bound to mouse D(b) major histocompatibility complex class I molecules, we show that ... Full text Link to item Cite

Signal sequence- and translation-independent mRNA localization to the endoplasmic reticulum.

Journal Article RNA · March 2008 The process of mRNA localization typically utilizes cis-targeting elements and trans-recognition factors to direct the compartmental organization of translationally suppressed mRNAs. mRNA localization to the endoplasmic reticulum (ER), in contrast, occurs ... Full text Link to item Cite

Divergent regulation of protein synthesis in the cytosol and endoplasmic reticulum compartments of mammalian cells.

Journal Article Mol Biol Cell · February 2008 In eukaryotic cells, mRNAs encoding signal sequence-bearing proteins undergo translation-dependent trafficking to the endoplasmic reticulum (ER), thereby restricting secretory and integral membrane protein synthesis to the ER compartment. However, recent s ... Full text Link to item Cite

Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells.

Journal Article Methods Mol Biol · 2008 All eukaryotic cells display a dramatic partitioning of mRNAs between the cytosol and endoplasmic reticulum (ER) compartments-mRNAs encoding secretory and integral membrane proteins are highly enriched on ER-bound ribosomes and mRNAs encoding cytoplasmic/n ... Full text Link to item Cite

In vitro and tissue culture methods for analysis of translation initiation on the endoplasmic reticulum.

Journal Article Methods Enzymol · 2007 For mRNAs encoding secretory and integral membrane proteins, translation initiation is thought to begin a process of mRNA localization where mRNA/ribosome/nascent chain complexes (RNCs) are trafficked from the cytosol compartment to the endoplasmic reticul ... Full text Link to item Cite

Interaction of TLR2 and TLR4 ligands with the N-terminal domain of Gp96 amplifies innate and adaptive immune responses.

Journal Article J Biol Chem · August 11, 2006 Featured Publication Activation of dendritic cells by ligands for Toll-like receptors (TLR) is a crucial event in the initiation of innate and adaptive immune responses. Several classes of TLR ligands have been identified that interact with distinct members of the TLR-family. ... Full text Link to item Cite

The DRiP hypothesis decennial: support, controversy, refinement and extension.

Journal Article Trends Immunol · August 2006 Featured Publication In 1996, to explain the rapid presentation of viral proteins to CD8+ T cells, it was proposed that peptides presented by MHC class I molecules derive from defective ribosomal products (DRiPs), presumed to be polypeptides arising from in-frame translation t ... Full text Link to item Cite

Synthesis of Hsp90 dimerization modulators.

Journal Article Bioorg Med Chem Lett · July 1, 2006 The synthesis and evaluation of several chemical modulators of heat shock protein 90 (Hsp90) dimerization is presented. These agents may represent useful tools to study the importance of N-terminal dimerization and also to determine subunit interface(s) in ... Full text Link to item Cite

Identification of novel quaternary domain interactions in the Hsp90 chaperone, GRP94.

Journal Article Protein Sci · June 2006 Featured Publication The structural basis for the coupling of ATP binding and hydrolysis to chaperone activity remains a central question in Hsp90 biology. By analogy to MutL, ATP binding to Hsp90 is thought to promote intramolecular N-terminal dimerization, yielding a molecul ... Full text Link to item Cite

mRNA translation is compartmentalized to the endoplasmic reticulum following physiological inhibition of cap-dependent translation.

Journal Article RNA · May 2006 Featured Publication Eukaryotic cells utilize a cycle of ribosome trafficking on the endoplasmic reticulum (ER) to partition mRNAs between the cytosol and ER compartments. In this process, ribosomes engaged in the synthesis of signal sequence-bearing proteins are trafficked to ... Full text Link to item Cite

Pathways for compartmentalizing protein synthesis in eukaryotic cells: the template-partitioning model.

Journal Article Biochem Cell Biol · December 2005 Featured Publication mRNAs encoding signal sequences are translated on endoplasmic reticulum (ER) -- bound ribosomes, whereas mRNAs encoding cytosolic proteins are translated on cytosolic ribosomes. The partitioning of mRNAs to the ER occurs by positive selection; cytosolic ri ... Full text Link to item Cite

Stable ribosome binding to the endoplasmic reticulum enables compartment-specific regulation of mRNA translation.

Journal Article Mol Biol Cell · December 2005 Featured Publication In eukaryotic cells, protein synthesis is compartmentalized; mRNAs encoding secretory/membrane proteins are translated on endoplasmic reticulum (ER)-bound ribosomes, whereas mRNAs encoding cytosolic proteins are translated on free ribosomes. mRNA partition ... Full text Link to item Cite

Enhancement of cancer radiation therapy by use of adenovirus-mediated secretable glucose-regulated protein 94/gp96 expression.

Journal Article Cancer Res · October 15, 2005 Tumor-derived glucose-regulated protein 94 (GRP94/gp96) has shown great promise as a tumor vaccine. However, current protein-based approaches require the availability of large quantities of tumor tissue, which are often not possible. In addition, the effic ... Full text Link to item Cite

HDAC6 regulates Hsp90 acetylation and chaperone-dependent activation of glucocorticoid receptor.

Journal Article Mol Cell · May 27, 2005 The molecular chaperone heat shock protein 90 (Hsp90) and its accessory cochaperones function by facilitating the structural maturation and complex assembly of client proteins, including steroid hormone receptors and selected kinases. By promoting the acti ... Full text Link to item Cite

Catheter-mediated subselective intracoronary gene delivery to the rabbit heart: introduction of a novel method.

Journal Article J Gene Med · May 2005 BACKGROUND: Recent studies suggest that gene therapy using replication-deficient adenoviruses will benefit treatment of cardiovascular diseases including heart failure. A persistent hurdle is the effective and reproducible delivery of a transgene to the my ... Full text Link to item Cite

Membrane topology mapping of vitamin K epoxide reductase by in vitro translation/cotranslocation.

Journal Article J Biol Chem · April 22, 2005 Vitamin K epoxide reductase (VKOR) catalyzes the conversion of vitamin K 2,3-epoxide into vitamin K in the vitamin K redox cycle. Recently, the gene encoding the catalytic subunit of VKOR was identified as a 163-amino acid integral membrane protein. In thi ... Full text Link to item Cite

Adenosine nucleotides and the regulation of GRP94-client protein interactions.

Journal Article Biochemistry · July 13, 2004 The molecular chaperone heat shock protein 90 (Hsp90) serves essential roles in the regulation of signaling protein function, trafficking, and turnover. Hsp90 function is intimately linked to intrinsic ATP binding and hydrolysis activities, the latter of w ... Full text Link to item Cite

Glucose-regulated protein 94/glycoprotein 96 elicits bystander activation of CD4+ T cell Th1 cytokine production in vivo.

Journal Article J Immunol · April 1, 2004 Glucose-regulated protein 94 (GRP94/gp96), the endoplasmic reticulum heat shock protein 90 paralog, elicits both innate and adaptive immune responses. Regarding the former, GRP94/gp96 stimulates APC cytokine expression and dendritic cell maturation. The ad ... Full text Link to item Cite

The messenger and the message: gp96 (GRP94)-peptide interactions in cellular immunity.

Journal Article Cell Stress Chaperones · 2004 Vaccination of mice with tumor-derived stress proteins, such as Hsp70 and gp96 (GRP94), can elicit antitumor immune responses, yielding a marked suppression of tumor growth and metastasis. The molecular basis for this response is proposed to reflect a pept ... Full text Link to item Cite

Structure of the N-terminal domain of GRP94. Basis for ligand specificity and regulation.

Journal Article J Biol Chem · November 28, 2003 GRP94, the endoplasmic reticulum (ER) paralog of the chaperone Hsp90, plays an essential role in the structural maturation or secretion of a subset of proteins destined for transport to the cell surface, such as the Toll-like receptors 2 and 4, and IgG, re ... Full text Link to item Cite

Scavenger receptor-A mediates gp96/GRP94 and calreticulin internalization by antigen-presenting cells.

Journal Article EMBO J · November 17, 2003 gp96 (GRP94) elicits antigen-presenting cell (APC) activation and can direct peptides into the cross- presentation pathways of APC. These responses arise through interactions of gp96 with Toll-like (APC activation) and endocytic (cross-presentation) recept ... Full text Link to item Cite

Partitioning and translation of mRNAs encoding soluble proteins on membrane-bound ribosomes.

Journal Article RNA · September 2003 In eukaryotic cells, it is generally accepted that protein synthesis is compartmentalized; soluble proteins are synthesized on free ribosomes, whereas secretory and membrane proteins are synthesized on endoplasmic reticulum (ER)-bound ribosomes. The partit ... Full text Link to item Cite

Come forth CD1d: Hsp110 in the regulation of intestinal epithelial CD1d expression.

Journal Article J Clin Invest · September 2003 CD1d, a nonclassical MHC class I-like molecule, is prominently expressed on intestinal epithelial cells and is thought to function in the regulation of intestinal intraepithelial lymphocyte activity. Hsp110, an abundant heat shock protein present in essent ... Full text Link to item Cite

GRP94/gp96 elicits ERK activation in murine macrophages. A role for endotoxin contamination in NF-kappa B activation and nitric oxide production.

Journal Article J Biol Chem · August 22, 2003 Vaccination of mice with GRP94/gp96, the endoplasmic reticulum Hsp90, elicits a variety of immune responses sufficient for tumor rejection and the suppression of metastatic tumor progression. Macrophages are a prominent GRP94/gp96 target, with GRP94/gp96 r ... Full text Link to item Cite

Re-evaluating the role of heat-shock protein-peptide interactions in tumour immunity.

Journal Article Nat Rev Immunol · May 2003 Early investigations into the immune surveillance of chemically-induced sarcomas led to two important concepts in tumour immunobiology: one, tumour rejection can be elicited by immune recognition of tumour antigens; and two, tumours express unique sets of ... Full text Link to item Cite

GRP94 (gp96) and GRP94 N-terminal geldanamycin binding domain elicit tissue nonrestricted tumor suppression.

Journal Article J Exp Med · December 2, 2002 Featured Publication In chemical carcinogenesis models, GRP94 (gp96) elicits tumor-specific protective immunity. The tumor specificity of this response is thought to reflect immune responses to GRP94-bound peptide antigens, the cohort of which uniquely identifies the GRP94 tis ... Full text Link to item Cite

Probing protein-protein interactions with chemical cross-linking and differential nano-LC-MS

Journal Article Proceedings 50th ASMS Conference on Mass Spectrometry and Allied Topics · December 1, 2002 Protein-protein interaction was investigated using chemical cross-linking and differential nano-LC-MS. The tryptic digestion mixtures were analyzed by nano-LC-MS. GGH-ecotin D137Y was used as a model protein and cross-linked by primary amine reactive cross ... Cite

A platform for compartmentalized protein synthesis: protein translation and translocation in the ER.

Journal Article Curr Opin Cell Biol · August 2002 Featured Publication Recent advances in the study of protein translocation across the membrane of the endoplasmic reticulum include insights into the mechanism of signal-sequence function. Biochemical and genetic studies have provided further evidence that lumenal proteins per ... Full text Link to item Cite

GRP94-associated enzymatic activities. Resolution by chromatographic fractionation.

Journal Article J Biol Chem · July 12, 2002 GRP94 (gp96), which performs established functions as a molecular chaperone and immune system modulator, has been reported to display a number of intrinsic enzymatic activities, including ATP hydrolysis, protein phosphorylation, and aminopeptidase. In obse ... Full text Link to item Cite

Endoplasmic reticulum-bound ribosomes reside in stable association with the translocon following termination of protein synthesis.

Journal Article J Biol Chem · June 28, 2002 Featured Publication In current views, translation-coupled ribosome binding to the endoplasmic reticulum (ER) membrane is transient, with association occurring via the signal recognition particle pathway and dissociation occurring upon the termination of protein synthesis. Rec ... Full text Link to item Cite

Transfer of GRP94(Gp96)-associated peptides onto endosomal MHC class I molecules.

Journal Article Traffic · May 2002 Featured Publication GRP94 (gp96)-associated peptides can elicit cellular immune responses, an activity thought to reflect the presence of a cell surface receptor (CD91) on antigen-presenting cells that mediates GRP94 internalization and trafficking to an amenable site for pep ... Full text Link to item Cite

Cutting edge: CD91-independent cross-presentation of GRP94(gp96)-associated peptides.

Journal Article J Immunol · May 1, 2002 Featured Publication GRP94(gp96) elicits CD8(+) T cell responses against its bound peptides, a process requiring access of its associated peptides into the MHC class I cross-presentation pathway of APCs. Entry into this pathway requires receptor-mediated endocytosis, and CD91 ... Full text Link to item Cite

Signal sequence function in the mammalian endoplasmic reticulum: A biological perspective

Journal Article Current Topics in Membranes · January 1, 2002 The discovery of the signal peptide, an amino-terminal protein sequence that specifies targeting of newly synthesized polypeptides to the endoplasmic reticulum (ER), stands as one of the most significant in cell biology. The signal peptide performs a targe ... Full text Cite

To find the road traveled to tumor immunity: the trafficking itineraries of molecular chaperones in antigen-presenting cells.

Journal Article Traffic · October 2001 Molecular chaperones, both endoplasmic reticulum and cytosol derived, have been identified as tumor rejection antigens; in animal models, they can elicit prophylactic and therapeutic immune responses against their tumor of origin. Chaperone immunogenic act ... Full text Link to item Cite

Virally induced lytic cell death elicits the release of immunogenic GRP94/gp96.

Journal Article J Biol Chem · June 15, 2001 Necrotic cell death yields the release of cellular components that can function in the initiation of cellular immune responses. Given the established capacity of the endoplasmic reticulum chaperone GRP94 (gp96) to elicit CD8(+) T cell activation, we have i ... Full text Link to item Cite

Ribosome exchange revisited: a mechanism for translation-coupled ribosome detachment from the ER membrane.

Journal Article Trends Cell Biol · March 2001 Featured Publication In current models, ribosome release from the endoplasmic reticulum (ER) is coupled to the termination of protein translation. Thus, coincident with termination, membrane-bound ribosomes dissociate into their component subunits and are released into the cyt ... Full text Link to item Cite

Sensitivity of mature Erbb2 to geldanamycin is conferred by its kinase domain and is mediated by the chaperone protein Hsp90.

Journal Article J Biol Chem · February 2, 2001 ErbB receptors are a family of ligand-activated tyrosine kinases that play a central role in proliferation, differentiation, and oncogenesis. ErbB2 is overexpressed in >25% of breast and ovarian cancers and is correlated with poor prognosis. Although ErbB2 ... Full text Link to item Cite

The fate of membrane-bound ribosomes following the termination of protein synthesis.

Journal Article J Biol Chem · October 27, 2000 Contemporary models for protein translocation in the mammalian endoplasmic reticulum (ER) identify the termination of protein synthesis as the signal for ribosome release from the ER membrane. We have utilized morphometric and biochemical methods to assess ... Full text Link to item Cite

Regulation of ribosome detachment from the mammalian endoplasmic reticulum membrane.

Journal Article J Biol Chem · October 27, 2000 Featured Publication In current models, protein translocation in the endoplasmic reticulum (ER) occurs in the context of two cycles, the signal recognition particle (SRP) cycle and the ribosome cycle. Both SRP and ribosomes bind to the ER membrane as a consequence of the targe ... Full text Link to item Cite

Chaperone-mediated cross-priming: a hitchhiker's guide to vesicle transport (review).

Journal Article Int J Mol Med · September 2000 The resident endoplasmic reticulum (ER) chaperone proteins GRP94 (gp96) and calreticulin can activate the immune system to slow or stop the progression of tumors by escorting tumor-derived peptides into the endogenous antigen presentation pathway of antige ... Full text Link to item Cite

A topological study of the human gamma-glutamyl carboxylase.

Journal Article Blood · August 1, 2000 gamma-Glutamyl carboxylase (GC), a polytopic membrane protein found in the endoplasmic reticulum (ER), catalyzes vitamin K-dependent posttranslational modification of glutamate to gamma-carboxyl glutamate. In an attempt to delineate the structure of this i ... Link to item Cite

Ligand interactions in the adenosine nucleotide-binding domain of the Hsp90 chaperone, GRP94. I. Evidence for allosteric regulation of ligand binding.

Journal Article J Biol Chem · July 28, 2000 Featured Publication X-ray crystallographic studies of the N-terminal domain of Hsp90 have identified an unconventional ATP binding fold, thereby inferring a role for ATP in the regulation of the Hsp90 activity. In this report, N-ethylcarboxamidoadenosine (NECA) was used to in ... Full text Link to item Cite

Ligand interactions in the adenosine nucleotide-binding domain of the Hsp90 chaperone, GRP94. II. Ligand-mediated activation of GRP94 molecular chaperone and peptide binding activity.

Journal Article J Biol Chem · July 28, 2000 Featured Publication The N-terminal domain of eukaryotic Hsp90 proteins contains a conserved adenosine nucleotide binding pocket that also serves as the binding site for the Hsp90 inhibitors geldanamycin and radicicol. Although this domain is essential for Hsp90 function, the ... Full text Link to item Cite

Role of chaperones in antigen processing.

Journal Article Immunol Invest · May 2000 Full text Link to item Cite

Ribosome-independent regulation of translocon composition and Sec61alpha conformation.

Journal Article J Biol Chem · January 21, 2000 In this study, the contributions of membrane-bound ribosomes to the regulation of endoplasmic reticulum translocon composition and Sec61alpha conformation were examined. Following solubilization of rough microsomes (RM) with digitonin, ribosomes co-sedimen ... Full text Link to item Cite

The immunological properties of endoplasmic reticulum chaperones: a conflict of interest?

Journal Article Essays Biochem · 2000 Featured Publication ER chaperones are abundant and highly conserved proteins that display both peptide binding and chaperone activity. Of the family of chaperones present in the mammalian ER, GRP94 and calreticulin are apparently unique in their ability to elicit CD8+ T-cell ... Full text Link to item Cite

Structural determinants for signal sequence function in the mammalian endoplasmic reticulum.

Journal Article J Biol Chem · December 17, 1999 Signal sequences function in protein targeting to and translocation across the endoplasmic reticulum membrane. To investigate the structural requirements for signal sequence function, chimeras of the Escherichia coli LamB signal peptide and prolactin were ... Full text Link to item Cite

Receptor mediated and fluid phase pathways for internalization of the ER Hsp90 chaperone GRP94 in murine macrophages.

Journal Article J Cell Sci · July 1999 Immunization of mice with GRP94, the endoplasmic reticulum (ER) Hsp90, elicits cytotoxic T lymphocyte (CTL) responses to chaperone-bound, source cell-derived peptides. Elicitation of a CTL response requires that GRP94-associated peptides be transferred ont ... Full text Link to item Cite

Calreticulin displays in vivo peptide-binding activity and can elicit CTL responses against bound peptides.

Journal Article J Immunol · June 1, 1999 Calreticulin is an endoplasmic reticulum (ER) chaperone that displays lectin activity and contributes to the folding pathways for nascent glycoproteins. Calreticulin also participates in the reactions yielding assembly of peptides onto nascent MHC class I ... Link to item Cite

Structural transitions accompanying the activation of peptide binding to the endoplasmic reticulum Hsp90 chaperone GRP94.

Journal Article Biochemistry · April 21, 1998 GRP94, the endoplasmic reticulum Hsp90 paralog, binds a diverse array of peptides, a subset of which are suitable for assembly onto nascent MHC class I molecules. At present, the mechanism, site, and regulation of peptide binding to GRP94 are unknown. Usin ... Full text Link to item Cite

Biochemical, cell biological and immunological issues surrounding the endoplasmic reticulum chaperone GRP94/gp96.

Journal Article Curr Opin Immunol · February 1998 The past year has born witness to compelling demonstrations of the utility of peptide complexes with glucose regulated protein 94 (GRP94, also known as gp96) in cancer immunotherapy. Insights into the structural basis of peptide binding to GRP94 have been ... Full text Link to item Cite

Regulation of the ribosome-membrane junction at early stages of presecretory protein translocation in the mammalian endoplasmic reticulum.

Journal Article J Cell Biol · December 29, 1997 A series of fusion protein constructs were designed to investigate the contribution of secretory nascent chains to regulation of the ribosome-membrane junction in the mammalian endoplasmic reticulum. As a component of these studies, the membrane topology o ... Full text Link to item Cite

Structure and folding of nascent polypeptide chains during protein translocation in the endoplasmic reticulum.

Journal Article J Biol Chem · July 4, 1997 To investigate the role of protein folding and chaperone-nascent chain interactions in translocation across the endoplasmic reticulum membrane, the translocation of wild type and mutant forms of preprolactin were studied in vivo and in vitro. The preprolac ... Full text Link to item Cite

Identification of a novel stage of ribosome/nascent chain association with the endoplasmic reticulum membrane.

Journal Article J Cell Biol · March 24, 1997 Protein translocation in the mammalian endoplasmic reticulum (ER) occurs cotranslationally and requires the binding of translationally active ribosomes to components of the ER membrane. Three candidate ribosome receptors, p180, p34, and Sec61p, have been i ... Full text Link to item Cite

Interaction of endoplasmic reticulum chaperone GRP94 with peptide substrates is adenine nucleotide-independent.

Journal Article J Biol Chem · February 21, 1997 GRP94, the endoplasmic reticulum paralog of hsp90, has recently been identified as a peptide and adenine nucleotide-binding protein. To determine if adenine nucleotides directly contribute to the regulation of GRP94 peptide binding activity, an in vitro pe ... Full text Link to item Cite

Membrane insertion, glycosylation, and oligomerization of inositol trisphosphate receptors in a cell-free translation system.

Journal Article J Biol Chem · January 17, 1997 In order to study the membrane topology, processing, and oligomerization of inositol trisphosphate receptor (IP3R) isoforms, we have utilized RNA templates encoding putative transmembrane domains to program a cell-free translation system of rabbit reticulo ... Full text Link to item Cite

Endoplasmic reticulum chaperone GRP94 subunit assembly is regulated through a defined oligomerization domain.

Journal Article Biochemistry · December 24, 1996 GRP94 is an abundant, resident glycoprotein of the mammalian endoplasmic reticulum lumen and member of the hsp90 family of molecular chaperones. To identify the structure/function relationships which define the molecular basis of GRP94 activity, we have pe ... Full text Link to item Cite

Biogenesis, cellular localization, and functional activation of the heat-stable enterotoxin receptor (guanylyl cyclase C).

Journal Article Biochemistry · August 20, 1996 Enterotoxigenic Escherichia coli elaborate a peptide called heat-stable enterotoxin (ST), which binds to and activates the intestinal ST receptor (STaR). STaR, also known as guanylyl cyclase C (GC-C), is a member of the transmembrane guanylyl cyclase recep ... Full text Link to item Cite

Purification and partial molecular characterization of GRP94, an ER resident chaperone.

Journal Article Protein Expr Purif · February 1996 GRP94 is a resident glycoprotein of the endoplasmic reticulum (ER) and a member of the hsp90 family of molecular chaperones. Current experimental evidence indicates that GRP94 functions in an as yet undefined manner in protein folding and assembly in the E ... Full text Link to item Cite

Protein translocation in the endoplasmic reticulum: The search for a unified molecular mechanism

Journal Article Seminars in Cell and Developmental Biology · January 1, 1996 The development of procedures for the reconstitution of protein translocation in the endoplasmic reticulum (ER) has allowed substantial new insights into the molecular mechanism of translocation. With these techniques, investigators are beginning to addres ... Full text Cite

Stage- and ribosome-specific alterations in nascent chain-Sec61p interactions accompany translocation across the ER membrane.

Journal Article J Cell Biol · May 1995 Near-neighbor interactions between translocating nascent chains and Sec61p were investigated by chemical cross-linking. At stages of translocation before signal sequence cleavage, nascent chains could be cross-linked to Sec61p at high (60-80%) efficiencies ... Full text Link to item Cite

Characterization of kinectin, a kinesin-binding protein: primary sequence and N-terminal topogenic signal analysis.

Journal Article Molecular biology of the cell · February 1995 Kinectin is a kinesin-binding protein (Toyoshima et al., 1992) that is required for kinesin-based motility (Kumar et al., 1995). A kinectin cDNA clone containing a 4.7-kilobase insert was isolated from an embryonic chick brain cDNA library by immunoscreeni ... Full text Cite

Lumenal proteins of the mammalian endoplasmic reticulum are required to complete protein translocation.

Journal Article Cell · June 4, 1993 The role of the lumenal contents (reticuloplasm) of the endoplasmic reticulum (ER) in protein translocation was determined by in vitro analysis. Depletion of the reticuloplasm from mammalian rough microsomes revealed two distinct stages of the translocatio ... Full text Link to item Cite

Membrane topology and biogenesis of eukaryotic signal peptidase.

Journal Article J Biol Chem · March 5, 1993 The signal peptidase complex (SPC) is a hetero-oligomeric membrane protein containing subunits of 12, 18, 21, 22/23, and 25 kDa. The 18- and 21-kDa subunits are mammalian homologs of SEC11 protein, which is necessary for signal peptide processing and cell ... Link to item Cite

The signal sequence receptor, unlike the signal recognition particle receptor, is not essential for protein translocation.

Journal Article J Cell Biol · April 1992 Detergent extracts of canine pancreas rough microsomal membranes were depleted of either the signal recognition particle receptor (SR), which mediates the signal recognition particle (SRP)-dependent targeting of the ribosome/nascent chain complex to the me ... Full text Link to item Cite

Biochemical fractionation and assembly of the membrane components that mediate nascent chain targeting and translocation.

Journal Article Cell · May 17, 1991 Fractionation of a microsomal detergent extract with ammonium sulfate allows separation of the signal recognition particle receptor (SR alpha), which is required for targeting of the nascent chain, from other microsomal proteins, such as signal peptidase, ... Full text Link to item Cite

Reconstitution of protein translocation from detergent-solubilized Escherichia coli inverted vesicles: PrlA protein-deficient vesicles efficiently translocate precursor proteins.

Journal Article Proc Natl Acad Sci U S A · March 1990 Proteoliposomes were reconstituted by detergent dialysis of a sodium cholate extract of inverted vesicles derived from Escherichia coli plasma membrane. The translocation of precursor proteins into reconstituted vesicles occurred at high efficiency and was ... Full text Link to item Cite

Assembly of translocation-competent proteoliposomes from detergent-solubilized rough microsomes.

Journal Article Cell · January 26, 1990 Canine pancreas rough microsomes were solubilized in a high salt buffer containing sodium cholate, a detergent extract prepared by high speed centrifugation, and vesicles were reconstituted from the extract by a detergent dialysis procedure. The reconstitu ... Full text Link to item Cite

Nascent secretory chain binding and translocation are distinct processes: differentiation by chemical alkylation.

Journal Article J Cell Biol · March 1989 We have investigated the effects of chemical alkylation of microsomal membranes on nascent chain binding and translocation. Assays were conducted using either full-length or truncated preprolactin transcripts in combination with a reconstituted membrane sy ... Full text Link to item Cite

Characteristics of GTP-mediated microsomal Ca2+ release.

Journal Article Biochim Biophys Acta · November 22, 1988 Guanosine triphosphate (GTP) can release Ca2+ and enhance responses to D-myo-inositol 1,4,5-trisphosphate (IP3) in crude liver microsomes in the presence of polyethylene glycol (PEG) (Dawson et al. (1986) Biochem. J. 234, 311-315). The mechanism of these r ... Full text Link to item Cite

GTP-mediated Ca2+ release in rough endoplasmic reticulum. Correlation with a GTP-sensitive increase in membrane permeability.

Journal Article Biochem J · December 15, 1987 Guanine nucleotides have been reported to stimulate reticular Ca2+ release. By using the structure-linked latency of microsomal mannose-6-phosphate phosphatase as an index of microsomal permeability [Arion, Ballas, Lange & Wallin (1976) J. Biol. Chem. 251, ... Full text Link to item Cite

Polyethylene glycol-stimulated microsomal GTP hydrolysis. Relationship to GTP-mediated Ca2+ release

Journal Article Journal of Pharmacology and Experimental Therapeutics · January 1, 1987 It has recently been observed that GTP mediates Ca2+ release from internal Ca2+ stores. In contrast to effects on permeabilized cells, GTP-dependent Ca2+ release in isolated microsomes requires the presence of polyethylene glycol (PEG). We have investigate ... Cite

Polyethylene glycol-stimulated microsomal GTP hydrolysis. Relationship to GTP-mediated Ca2+ release.

Journal Article FEBS Lett · December 15, 1986 It has recently been observed that GTP mediates Ca2+ release from internal Ca2+ stores. In contrast to effects on permeabilized cells, GTP-dependent Ca2+ release in isolated microsomes requires the presence of polyethylene glycol (PEG). We have investigate ... Full text Link to item Cite

Cyclosporine augments receptor-mediated cellular Ca2+ fluxes in isolated hepatocytes.

Journal Article J Biol Chem · November 5, 1985 The immunosuppressive agent, cyclosporine, has been found to augment receptor-stimulated calcium fluxes in isolated hepatocytes. After treatment of Quin 2-loaded hepatocytes with cyclosporine, both the amplitude and duration of the vasopressin-induced rise ... Link to item Cite

Spermine. A regulator of mitochondrial calcium cycling.

Journal Article J Biol Chem · November 10, 1984 Steady-state free Ca2+ concentrations have been measured with a Ca2+ electrode using suspensions of isolated rat liver mitochondria or saponin-treated hepatocytes. Mitochondria, when incubated in the presence of Mg2+ and MgATP2-, maintain a steady-state pC ... Link to item Cite

High-performance liquid chromatographic separation and quantification of alanopine and strombine in crude tissue extracts.

Journal Article Anal Biochem · June 1984 A method for the separation and quantification of the levels of alanopine and strombine in neutralized, perchloric acid extracts of tissues of marine invertebrates is presented. The method is based on high-performance liquid chromatographic (HPLC) separati ... Full text Link to item Cite

Effects of spermine on Ca2+-cycling in rat liver mitochondria

Journal Article Federation Proceedings · January 1, 1984 Cite

Partial purification and characterization of a strombine dehydrogenase from the adductor muscle of the mussel Modiolus squamosus

Journal Article Comparative Biochemistry and Physiology -- Part B: Biochemistry and · January 1, 1984 1. 1. Strombine dehydrogenase from the adductor muscle of the marine mussel Modiolus squamosus was partially purified by ammonium sulfate fractionation, gel filtration and chromatography on Procion Red Agarose. 2. 2. The enzyme showed relatively broad amin ... Full text Cite