Scholars@Duke
Michael C. Fitzgerald

Michael C. Fitzgerald

Professor of Chemistry
Chemistry
michael.c.fitzgerald@duke.edu
Box 90346, Durham, NC 27708-0346
3222 French Science Center, 124 Science Drive, Durham, NC 27708

Selected Publications

Discovery of Host-Directed Small Molecules with Broad Anti-Leishmanial Efficacy.

Journal Article bioRxiv · November 4, 2025 Leishmaniasis, a neglected tropical disease affecting nearly 10% of the global population, suffers from limited therapeutic options and rising drug resistance. To address this, we developed 343 analogs of AR-12, a compound that has previously illustrated h ... Full text Link to item Cite

Leveraging Vulnerabilities in Copper Trafficking for Synergistic Antifungal Activity.

Journal Article ACS chemical biology · November 2025 Candida albicans is an opportunistic fungal pathogen that causes millions of infections per year, for which more efficacious treatments are needed. Observations that azole antifungals incite C. albicans to adjust a variety of metal-dependent ... Full text Cite

Copper Activates a Redox Switch to Reversibly Inhibit Glyceraldehyde-3-Phosphate Dehydrogenase.

Journal Article Biochemistry · November 2025 Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), one of the most conserved proteins across all kingdoms of life, has a multitude of moonlighting functions beyond its enzymatic role in glycolysis. Metal binding to GAPDH has previously been reported to inhi ... Full text Cite

Extrinsic and intrinsic factors affect copper-induced protein precipitation across eukaryotic and prokaryotic proteomes.

Journal Article Protein science : a publication of the Protein Society · June 2025 The susceptibility of a protein to aggregation upon exposure to copper ions (Cu) has been recognized as a contributor to Cu-induced cellular dysfunction and toxicity. Different cell types succumb to Cu to varying degrees, indicating innate differences betw ... Full text Cite

Covalent inhibition of <i>Plasmodium falciparum</i> Ubc13 impairs global protein synthesis.

Journal Article iScience · June 2025 The ubiquitin-conjugating enzyme 13 (Ubc13) has an essential function and putative role in artemisinin activity against Plasmodium falciparum. Ubc13 conjugates lysine 63-linked ubiquitin (K63-Ub) to proteins, but the role of this modification in ... Full text Cite

Top-Down Stability of Proteins from Rates of Oxidation (TD-SPROX) Approach for Measuring Proteoform-Specific Folding Stability.

Journal Article Analytical chemistry · December 2024 The crucial roles of proteoforms in biological processes and disease mechanisms have been increasingly recognized. However, the rate at which new proteoforms are being discovered using top-down proteomics has far outpaced the rate at which the functional s ... Full text Cite

Analysis of Brain Protein Stability Changes in a Mouse Model of Alzheimer's Disease.

Journal Article Journal of proteome research · October 2024 The stability of proteins from rates of oxidation (SPROX), thermal proteome profiling (TPP), and limited proteolysis (LiP) techniques were used to profile the stability of ∼2500 proteins in hippocampus tissue cell lysates from 2- and 8-months-old wild-type ... Full text Cite

Selective targeting of Plasmodium falciparum Hsp90 disrupts the 26S proteasome.

Journal Article Cell Chem Biol · April 18, 2024 The molecular chaperone heat shock protein 90 (Hsp90) has an essential but largely undefined role in maintaining proteostasis in Plasmodium falciparum, the most lethal malaria parasite. Herein, we identify BX-2819 and XL888 as potent P. falciparum (Pf)Hsp9 ... Full text Link to item Cite

Stability-Based Proteomics for Investigation of Structured RNA-Protein Interactions.

Journal Article Analytical chemistry · February 2024 RNA-protein interactions are essential to RNA function throughout biology. Identifying the protein interactions associated with a specific RNA, however, is currently hindered by the need for RNA labeling or costly tiling-based approaches. Conventional stra ... Full text Cite

Protein Folding Stability Profiling of Colorectal Cancer Chemoresistance Identifies Functionally Relevant Biomarkers.

Journal Article Journal of proteome research · June 2023 Reported here is the application of three protein folding stability profiling techniques (including the stability of proteins from rates of oxidation, thermal protein profiling, and limited proteolysis approaches) to identify differentially stabilized prot ... Full text Cite

Comparative Analysis of Protein Folding Stability-Based Profiling Methods for Characterization of Biological Phenotypes.

Journal Article Journal of the American Society for Mass Spectrometry · March 2023 Recently, a new suite of mass spectrometry-based proteomic methods has been developed that enables evaluation of protein folding stability on the proteomic scale. These methods utilize chemical and thermal denaturation approaches (SPROX and TPP, respective ... Full text Cite

Analysis of copper-induced protein precipitation across the E. coli proteome.

Journal Article Metallomics : integrated biometal science · January 2023 Metal cations have been exploited for their precipitation properties in a wide variety of studies, ranging from differentiating proteins from serum and blood to identifying the protein targets of drugs. Despite widespread recognition of this phenomenon, th ... Full text Cite

Discovery of the Xenon-Protein Interactome Using Large-Scale Measurements of Protein Folding and Stability.

Journal Article Journal of the American Chemical Society · March 2022 The intermolecular interactions of noble gases in biological systems are associated with numerous biochemical responses, including apoptosis, inflammation, anesthesia, analgesia, and neuroprotection. The molecular modes of action underlying these responses ... Full text Cite

Chemoproteomic-enabled characterization of small GTPase Rab1a as a target of an N-arylbenzimidazole ligand's rescue of Parkinson's-associated cell toxicity.

Journal Article RSC chemical biology · January 2022 The development of phenotypic models of Parkinson's disease (PD) has enabled screening and identification of phenotypically active small molecules that restore complex biological pathways affected by PD toxicity. While these phenotypic screening platforms ... Full text Cite

Analysis of Brain Protein Stability Changes in Mouse Models of Normal Aging and α-Synucleinopathy Reveals Age- and Disease-Related Differences.

Journal Article Journal of proteome research · November 2021 Here, we utilize the stability of proteins from rates of oxidation (SPROX) technique, to profile the thermodynamic stabilities of proteins in brain tissue cell lysates from Huα-Syn(A53T) transgenic mice at three time points including at 1 month (n = ... Full text Cite

Chemoproteomics for Plasmodium Parasite Drug Target Discovery.

Journal Article Chembiochem : a European journal of chemical biology · August 2021 Emerging Plasmodium parasite drug resistance is threatening progress towards malaria control and elimination. While recent efforts in cell-based, high-throughput drug screening have produced first-in-class drugs with promising activities against different ... Full text Cite

Protein Folding Stability Changes Across the Proteome Reveal Targets of Cu Toxicity in E. coli.

Journal Article ACS chemical biology · January 2021 The ability of metal ionophores to induce cellular metal hyperaccumulation endows them with potent antimicrobial activity; however, the targets and mechanisms behind these outcomes are not well understood. This work describes the first utilization of prote ... Full text Cite

Plasmodium chaperonin TRiC/CCT identified as a target of the antihistamine clemastine using parallel chemoproteomic strategy.

Journal Article Proceedings of the National Academy of Sciences of the United States of America · March 2020 Featured Publication The antihistamine clemastine inhibits multiple stages of the Plasmodium parasite that causes malaria, but the molecular targets responsible for its parasite inhibition were unknown. Here, we applied parallel chemoproteomic platforms to discover the ... Full text Cite

Comparative Analysis of Mass-Spectrometry-Based Proteomic Methods for Protein Target Discovery Using a One-Pot Approach.

Journal Article Journal of the American Society for Mass Spectrometry · February 2020 Featured Publication Recently, several mass-spectrometry- and protein-denaturation-based proteomic methods have been developed to facilitate protein target discovery efforts in drug mode-of-action studies. These methods, which include the stability of proteins from rates of ox ... Full text Cite

Chemo-Selection Strategy for Limited Proteolysis Experiments on the Proteomic Scale.

Journal Article Analytical chemistry · December 2018 Described here is a chemo-selective enrichment strategy, termed the semitryptic peptide enrichment strategy for proteolysis procedures (STEPP), to isolate the semitryptic peptides generated in mass spectrometry-based proteome-wide applications of limited p ... Full text Cite

Proteome-Wide Structural Biology: An Emerging Field for the Structural Analysis of Proteins on the Proteomic Scale.

Journal Article Journal of proteome research · November 2018 Over the past decade, a suite of new mass-spectrometry-based proteomics methods has been developed that now enables the conformational properties of proteins and protein-ligand complexes to be studied in complex biological mixtures, from cell lysates to in ... Full text Cite

Chemical Denaturation and Protein Precipitation Approach for Discovery and Quantitation of Protein-Drug Interactions.

Journal Article Analytical chemistry · August 2018 Described here is a mass spectrometry-based proteomics approach for the large-scale analysis of protein-drug interactions. The approach involves the evaluation of ligand-induced protein folding free energy changes (ΔΔ Gf) using chemical denatura ... Full text Cite

Proteome-Wide Characterization of Phosphorylation-Induced Conformational Changes in Breast Cancer.

Journal Article Journal of proteome research · March 2018 Because of the close link between protein function and protein folding stability, knowledge about phosphorylation-induced protein folding stability changes can lead to a better understanding of the functional effects of protein phosphorylation. Here, the s ... Full text Cite

Discovery of Tamoxifen and N-Desmethyl Tamoxifen Protein Targets in MCF-7 Cells Using Large-Scale Protein Folding and Stability Measurements.

Journal Article Journal of proteome research · November 2017 The proteins in an MCF-7 cell line were probed for tamoxifen (TAM) and n-desmethyl tamoxifen (NDT) induced stability changes using the Stability of Proteins from Rates of Oxidation (SPROX) technique in combination with two different quantitative proteomics ... Full text Cite

Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using SILAC-SPROX.

Journal Article Journal of proteome research · September 2017 Proteomic methods for disease state characterization and biomarker discovery have traditionally utilized quantitative mass spectrometry methods to identify proteins with altered expression levels in disease states. Here we report on the large-scale use of ... Full text Cite

Pathogenic Mutations Induce Partial Structural Changes in the Native β-Sheet Structure of Transthyretin and Accelerate Aggregation.

Journal Article Biochemistry · September 2017 Amyloid formation of natively folded proteins involves global and/or local unfolding of the native state to form aggregation-prone intermediates. Here we report solid-state nuclear magnetic resonance (NMR) structural studies of amyloid derived from wild-ty ... Full text Cite

Proteases of Dermatophagoides pteronyssinus.

Journal Article International journal of molecular sciences · June 2017 Since the discovery that Der p 1 is a cysteine protease, the role of proteolytic activity in allergic sensitization has been explored. There are many allergens with proteolytic activity; however, exposure from dust mites is not limited to allergens. In thi ... Full text Cite

Discovery of Age-Related Protein Folding Stability Differences in the Mouse Brain Proteome.

Journal Article Journal of proteome research · December 2016 Described here is the application of thermodynamic stability measurements to study age-related differences in the folding and stability of proteins in a rodent model of aging. Thermodynamic stability profiles were generated for 809 proteins in brain cell l ... Full text Cite

Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using Limited Proteolysis.

Journal Article Journal of proteome research · December 2016 Conformational changes in proteins can lead to disease. Thus, methods for identifying conformational changes in proteins can further improve our understanding and facilitate detection of disease states. Here we combine limited proteolysis (LiP) with Stable ... Full text Cite

Targeted Mass Spectrometry-Based Approach for Protein-Ligand Binding Analyses in Complex Biological Mixtures Using a Phenacyl Bromide Modification Strategy.

Journal Article Analytical chemistry · November 2016 The characterization of protein folding stability changes on the proteomic scale is useful for protein-target discovery and for the characterization of biological states. The Stability of Proteins from Rates of Oxidation (SPROX) technique is one of several ... Full text Cite

Thermodynamic Analysis of the Geldanamycin-Hsp90 Interaction in a Whole Cell Lysate Using a Mass Spectrometry-Based Proteomics Approach.

Journal Article Journal of the American Society for Mass Spectrometry · October 2016 Geldanamycin is a natural product with well-established and potent anti-cancer activities. Heat shock protein 90 (Hsp90) is the known target of geldanamycin, which directly binds to Hsp90's N-terminal ATP binding domain and inhibits Hsp90's ATPase activity ... Full text Cite

Discovery of Manassantin A Protein Targets Using Large-Scale Protein Folding and Stability Measurements.

Journal Article J Proteome Res · August 5, 2016 Manassantin A is a natural product that has been shown to have anticancer activity in cell-based assays, but has a largely unknown mode-of-action. Described here is the use of two different energetics-based approaches to identify protein targets of manassa ... Full text Link to item Cite

Characterization of the Saccharomyces cerevisiae ATP-Interactome using the iTRAQ-SPROX Technique.

Journal Article Journal of the American Society for Mass Spectrometry · February 2016 The stability of proteins from rates of oxidation (SPROX) technique was used in combination with an isobaric mass tagging strategy to identify adenosine triphosphate (ATP) interacting proteins in the Saccharomyces cerevisiae proteome. The SPROX methodology ... Full text Cite

Synthesis and Biological Evaluation of Manassantin Analogues for Hypoxia-Inducible Factor 1α Inhibition.

Journal Article J Med Chem · October 8, 2015 To cope with hypoxia, tumor cells have developed a number of adaptive mechanisms mediated by hypoxia-inducible factor 1 (HIF-1) to promote angiogenesis and cell survival. Due to significant roles of HIF-1 in the initiation, progression, metastasis, and res ... Full text Link to item Cite

Global analysis of protein folding thermodynamics for disease state characterization.

Journal Article Journal of proteome research · May 2015 Current methods for the large-scale characterization of disease states generally rely on the analysis of gene and/or protein expression levels. These existing methods fail to detect proteins with disease-related functions and unaltered expression levels. H ... Full text Cite

SILAC-pulse proteolysis: A mass spectrometry-based method for discovery and cross-validation in proteome-wide studies of ligand binding.

Journal Article Journal of the American Society for Mass Spectrometry · December 2014 Reported here is the use of stable isotope labeling with amino acids in cell culture (SILAC) and pulse proteolysis (PP) for detection and quantitation of protein-ligand binding interactions on the proteomic scale. The incorporation of SILAC into PP enables ... Full text Cite

StableIsotope Labeling with Amino Acids in Cell Culture (SILAC)-based strategy for proteome-wide thermodynamic analysis of protein-ligand binding interactions.

Journal Article Molecular & cellular proteomics : MCP · July 2014 Described here is a quantitative mass spectrometry-based proteomics method for the large-scale thermodynamic analysis of protein-ligand binding interactions. The methodology utilizes a chemical modification strategy termed, Stability of Proteins from Rates ... Full text Cite

Thermodynamic analysis of protein folding and stability using a tryptophan modification protocol.

Journal Article Analytical chemistry · July 2014 Described here is the development of a mass spectrometry-based covalent labeling protocol that utilizes the reaction of dimethyl(2-hydroxy-5-nitrobenzyl)sulfonium bromide (HNSB) with tryptophan (Trp) residues to measure protein folding free energies (ΔG(f) ... Full text Cite

Energetics-based methods for protein folding and stability measurements.

Journal Article Annual review of analytical chemistry (Palo Alto, Calif.) · January 2014 Over the past 15 years, a series of energetics-based techniques have been developed for the thermodynamic analysis of protein folding and stability. These techniques include Stability of Unpurified Proteins from Rates of amide H/D Exchange (SUPREX), pulse ... Full text Cite

False-positive rate determination of protein target discovery using a covalent modification- and mass spectrometry-based proteomics platform.

Journal Article Journal of the American Society for Mass Spectrometry · January 2014 Detection and quantitation of protein-ligand binding interactions is important in many areas of biological research. Stability of proteins from rates of oxidation (SPROX) is an energetics-based technique for identifying the proteins targets of ligands in c ... Full text Cite

Thermodynamic analysis of protein-ligand binding interactions in complex biological mixtures using the stability of proteins from rates of oxidation.

Journal Article Nature protocols · January 2013 The detection and quantification of protein-ligand binding interactions is crucial in a number of different areas of biochemical research from fundamental studies of biological processes to drug discovery efforts. Described here is a protocol that can be u ... Full text Cite

Evidence of Fe3+ interaction with the plug domain of the outer membrane transferrin receptor protein of Neisseria gonorrhoeae: implications for Fe transport.

Journal Article Metallomics : integrated biometal science · April 2012 Neisseria gonorrhoeae is an obligate pathogen that hijacks iron from the human iron transport protein, holo-transferrin (Fe(2)-Tf), by expressing TonB-dependent outer membrane receptor proteins, TbpA and TbpB. Homologous to other TonB-dependent outer membr ... Full text Cite

Order out of disorder: working cycle of an intrinsically unfolded chaperone.

Journal Article Cell · March 2012 The redox-regulated chaperone Hsp33 protects organisms against oxidative stress that leads to protein unfolding. Activation of Hsp33 is triggered by the oxidative unfolding of its own redox-sensor domain, making Hsp33 a member of a recently discovered clas ... Full text Cite

Slow histidine H/D exchange protocol for thermodynamic analysis of protein folding and stability using mass spectrometry.

Journal Article Analytical chemistry · February 2012 Described here is a mass spectrometry-based protocol to study the thermodynamic stability of proteins and protein-ligand complexes using the chemical denaturant dependence of the slow H/D exchange reaction of the imidazole C(2) proton in histidine side cha ... Full text Cite

Thermodynamic analysis of protein-ligand interactions in complex biological mixtures using a shotgun proteomics approach.

Journal Article Journal of proteome research · November 2011 Shotgun proteomics protocols are widely used for the identification and/or quantitation of proteins in complex biological samples. Described here is a shotgun proteomics protocol that can be used to identify the protein targets of biologically relevant lig ... Full text Cite

Mass spectrometry- and lysine amidination-based protocol for thermodynamic analysis of protein folding and ligand binding interactions.

Journal Article Analytical chemistry · May 2011 Described here is a mass spectrometry-based covalent labeling protocol that utilizes the amine reactive reagent, s-methyl thioacetimidate (SMTA), to study the chemical denaturant-induced equilibrium unfolding/refolding properties of proteins and protein-li ... Full text Cite

Stable isotope labeling strategy for protein-ligand binding analysis in multi-component protein mixtures.

Journal Article Journal of the American Society for Mass Spectrometry · March 2011 Described here is a stable isotope labeling protocol that can be used with a chemical modification- and mass spectrometry-based protein-ligand binding assay for detecting and quantifying both the direct and indirect binding events that result from protein- ... Full text Cite

Xu, Y., Falk, I.N., Hallen, M.A., and Fitzgerald, M.C.

Journal Article Analytical Chemistry · 2011 Cite

Discovery of novel cyclophilin A ligands using an H/D exchange- and mass spectrometry-based strategy.

Journal Article Journal of biomolecular screening · October 2010 Cyclophilin A (CypA) is an overexpressed protein in lung cancer tumors and as a result is a potential therapeutic and diagnostic target. Described here is use of an H/D exchange- and a matrix assisted laser desorption/ionization (MALDI) mass spectrometry-b ... Full text Cite

Total synthesis, assignment of the absolute stereochemistry, and structure-activity relationship studies of subglutinols A and B.

Journal Article Chemistry, an Asian journal · August 2010 Immunosuppressive drugs are used to prevent rejection of transplanted organs and treat autoimmune diseases. Clinically approved immunosuppressive drugs possess undesirable side effects, including acute neurological toxicity, chronic nephrotoxicity, and ost ... Full text Cite

Mass spectrometry-based thermal shift assay for protein-ligand binding analysis.

Journal Article Anal Chem · July 1, 2010 Described here is a mass spectrometry-based screening assay for the detection of protein-ligand binding interactions in multicomponent protein mixtures. The assay utilizes an oxidation labeling protocol that involves using hydrogen peroxide to selectively ... Full text Open Access Link to item Cite

Quantitative proteomics approach for identifying protein-drug interactions in complex mixtures using protein stability measurements.

Journal Article Proceedings of the National Academy of Sciences of the United States of America · May 2010 Knowledge about the protein targets of therapeutic agents is critical for understanding drug mode of action. Described here is a mass spectrometry-based proteomics method for identifying the protein target(s) of drug molecules that is potentially applicabl ... Full text Cite

Thermodynamic analysis of a molecular chaperone binding to unfolded protein substrates.

Journal Article Biochemistry · February 2010 Molecular chaperones are a highly diverse group of proteins that recognize and bind unfolded proteins to facilitate protein folding and prevent nonspecific protein aggregation. The mechanisms by which chaperones bind their protein substrates have been stud ... Full text Open Access Cite

A protein-ligand binding assay with proteomic potential

Conference ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY · August 16, 2009 Link to item Cite

Hydrogen/deuterium exchange- and protease digestion-based screening assay for protein-ligand binding detection.

Journal Article Analytical chemistry · August 2009 A protease digestion strategy was incorporated into single-point stability of unpurified proteins from rates of H/D exchange (SUPREX), which is a hydrogen/deuterium (H/D) exchange- and mass spectrometry-based assay for the detection of protein-ligand bindi ... Full text Cite

Painting proteins with covalent labels: what's in the picture?

Journal Article Journal of the American Society for Mass Spectrometry · June 2009 Knowledge about the structural and biophysical properties of proteins when they are free in solution and/or in complexes with other molecules is essential for understanding the biological processes that proteins regulate. Such knowledge is also important t ... Full text Cite

Hijacking transferrin bound iron: protein-receptor interactions involved in iron transport in N. gonorrhoeae.

Journal Article Metallomics : integrated biometal science · January 2009 Neisseria gonorrhoeae has the capacity to acquire iron from its human host by removing this essential nutrient from serum transferrin. The transferrin binding proteins, TbpA and TbpB constitute the outer membrane receptor complex responsible for binding tr ... Full text Cite

Chapter 6 Thermodynamic Analysis of Protein Folding and Ligand Binding by SUPREX

Journal Article Comprehensive Analytical Chemistry · December 1, 2008 Full text Cite

In vivo and in vitro examination of stability of primary hyperoxaluria-associated human alanine:glyoxylate aminotransferase.

Journal Article The Journal of biological chemistry · November 2008 Primary hyperoxaluria type I is a severe kidney stone disease caused by mutations in the protein alanine:glyoxylate aminotransferase (AGT). Many patients have mutations in AGT that are not deleterious alone but act synergistically with a common minor allel ... Full text Cite

Throughput and efficiency of a mass spectrometry-based screening assay for protein-ligand binding detection.

Journal Article Journal of the American Society for Mass Spectrometry · September 2008 An H/D exchange- and MALDI mass spectrometry-based screening assay was applied to search for novel ligands that bind to cyclophilin A, a potential therapeutic and diagnostic target in lung cancer. The assay is based on stability of unpurified proteins from ... Full text Cite

CHED 160-Thermodynamic analysis of proteins in the T7 bacteriophage proteome

Conference ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY · August 17, 2008 Link to item Cite

Ga3+ as a mechanistic probe in Fe3+ transport: characterization of Ga3+ interaction with FbpA.

Journal Article Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry · August 2008 The obligate human pathogens Haemophilus influenzae, Neisseria gonorrhoeae, and N. meningitidis utilize a highly conserved, three-protein ATP-binding cassette transporter (FbpABC) to shuttle free Fe(3+) from the periplasm and across the cytoplasmic membran ... Full text Cite

Thermodynamic analysis of protein stability and ligand binding using a chemical modification- and mass spectrometry-based strategy.

Journal Article Analytical chemistry · June 2008 Described here is a new technique, termed SPROX (stability of proteins from rates of oxidation), that can be used to measure the thermodynamic stability of proteins and protein-ligand complexes. SPROX utilizes hydrogen peroxide in the presence of increasin ... Full text Cite

Ex vivo analysis of synergistic anion binding to FbpA in Gram-negative bacteria.

Journal Article Biochemistry · April 2008 Ferric binding protein, FbpA, is a member of the transferrin superfamily whose function is to move an essential nutrient, iron, across the periplasm and into the cytosol through formation of a ternary complex containing Fe (3+) and a synergistic anion, X. ... Full text Cite

H/D exchange and mass spectrometry-based method for biophysical analysis of multidomain proteins at the domain level.

Journal Article Analytical chemistry · November 2007 A protocol was developed to characterize the domain-specific thermodynamic stabilities of multidomain proteins using SUPREX (Stability of Unpurified Proteins from Rates of H/D Exchange). The protocol incorporates a protease digestion step into the conventi ... Full text Cite

H/D exchange- and mass spectrometry-based strategy for the thermodynamic analysis of protein-ligand binding.

Journal Article Analytical chemistry · August 2007 The equilibrium unfolding properties of four model protein systems were characterized using SUPREX (stability of unpurified proteins from rates of H/D exchange). SUPREX is an H/D exchange- and mass spectrometry-based technique for measuring the free energy ... Full text Cite

Structural and thermodynamic characterization of a cytoplasmic dynein light chain-intermediate chain complex.

Journal Article Proceedings of the National Academy of Sciences of the United States of America · June 2007 Cytoplasmic dynein is a microtubule-based motor protein complex that plays important roles in a wide range of fundamental cellular processes, including vesicular transport, mitosis, and cell migration. A single major form of cytoplasmic dynein associates w ... Full text Cite

Mutational analysis of active site residues in the Staphylococcus aureus transpeptidase SrtA.

Journal Article Biochemistry · June 2007 In Staphylococcus aureus, virulence and colonization-associated surface proteins are covalently anchored to the cell wall by the transpeptidase Sortase A (SrtA). In order to better understand the contribution of specific active site residues to substrate r ... Full text Cite

Accuracy of SUPREX (stability of unpurified proteins from rates of H/D exchange) and MALDI mass spectrometry-derived protein unfolding free energies determined under non-EX2 exchange conditions.

Journal Article Journal of the American Society for Mass Spectrometry · November 2006 Described here is the impact of so-called non-EX2 exchange behavior on the accuracy of protein unfolding free energies (i.e., DeltaG u values) and m values (i.e.,-deltaDeltaG u/delta[denaturant] values) determined by an H/D exchange and mass spectrometry-b ... Full text Cite

A mass spectrometry-based probe of equilibrium intermediates in protein-folding reactions.

Journal Article Biochemistry · October 2006 Described here is a mass spectrometry- and H/D exchange-based approach for the detection of equilibrium intermediate state(s) in protein-folding reactions. The approach utilizes the stability of unpurified proteins from rates of H/D exchange (SUPREX) techn ... Full text Cite

Direct analysis of backbone-backbone hydrogen bond formation in protein folding transition states.

Journal Article Journal of molecular biology · October 2006 Here we investigate the role of backbone-backbone hydrogen bonding interactions in stabilizing the protein folding transition states of two model protein systems, the B1 domain of protein L (ProtL) and the P22 Arc repressor. A backbone modified analogue of ... Full text Cite

Total chemical synthesis of the B1 domain of protein L from Peptostreptococcus magnus.

Journal Article Bioorganic chemistry · June 2006 Reported here is a native chemical ligation strategy for the total chemical synthesis of the B1 domain of protein L. A synthetic construct of this 76 amino acid protein domain was prepared by the chemoselective ligation of two unprotected polypeptide fragm ... Full text Cite

Conserved thermodynamic contributions of backbone hydrogen bonds in a protein fold.

Journal Article Proceedings of the National Academy of Sciences of the United States of America · February 2006 Backbone-backbone hydrogen-bonding interactions are a ubiquitous and highly conserved structural feature of proteins that adopt the same fold (i.e., have the same overall backbone topology). This work addresses the question of whether or not this structura ... Full text Cite

Theoretical and experimental determination on two substrates turned over by 4-oxalocrotonate tautomerase.

Journal Article The journal of physical chemistry. A · January 2006 Quantum mechanical/molecular mechanical (QM/MM) calculations and experimental kinetic studies have been performed on 4-oxalocrotonate tautomerase (4OT) for two different substrates, 2-hydroxymuconate (2HM) and 2-oxo-4-hexenedioate (2o4hex). Potential (delt ... Full text Cite

SUPREX analysis of the thermodynamics of synergistic anion binding by ferric binding protein

Conference ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY · March 13, 2005 Link to item Cite

Thermodynamic analysis of subunit interactions in Escherichia coli molybdopterin synthase.

Journal Article Biochemistry · February 2005 The molybdopterin (MPT) synthase complex in Escherichia coli consists of two MoaE subunits and two MoaD subunits in a heterotetrameric structure with the two MoaE subunits forming a central dimer. Each MoaD subunit binds to a single MoaE molecule to form t ... Full text Cite

The Thermodynamics of Backbone-Backbone Hydrogen Bonds are Conserved ina Protein Fold

Journal Article Proceedings of the National Academy of Sciences, USA · 2005 Cite

Characterising phase variations in MALDI-TOF data and correcting them by peak alignment.

Journal Article Cancer informatics · January 2005 The use of MALDI-TOF mass spectrometry as a means of analyzing the proteome has been evaluated extensively in recent years. One of the limitations of this technique that has impeded the development of robust data analysis algorithms is the variability in t ... Open Access Cite

Protocol for the thermodynamic analysis of some proteins using an H/D exchange- and mass spectrometry-based technique.

Journal Article Analytical chemistry · January 2005 SUPREX (stability of unpurified proteins from rates of H/D exchange) is a new H/D exchange- and mass spectrometry-based technique for the measurement of protein folding free energies (i.e., DeltaG values) and protein folding m values (i.e., deltaDeltaG/del ... Full text Cite

SUPREX (Stability of Unpurified Proteins from Rates of H/D Exchange) analysis of the thermodynamics of synergistic anion binding by ferric-binding protein (FbpA), a bacterial transferrin.

Journal Article Biochemistry · December 2004 SUPREX (stability of unpurified proteins from rates of H/D exchange) is a H/D exchange- and matrix-assisted laser desorption/ionization (MALDI)-based technique for characterizing the equilibrium unfolding/refolding properties of proteins and protein-ligand ... Full text Cite

Translating biomarkers into clinical practice: prognostic implications of cyclophilin A and macrophage migratory inhibitory factor identified from protein expression profiles in non-small cell lung cancer.

Journal Article Lung Cancer · December 2004 Biomarkers have the potential to significantly change diagnostic strategies and influence therapeutic management. We developed a MALDI-TOF protein expression profiling platform for biomarker discovery and a proof-of-principle study identified two proteins, ... Full text Link to item Cite

Analysis of protein folding and function using backbone modified proteins.

Journal Article Bioorganic chemistry · October 2004 With the recent development of chemical and biological methods to introduce backbone modifications into the polypeptide chains of proteins, there have been a growing number of site-directed mutagenesis experiments focused on understanding the role of the p ... Full text Cite

Thermodynamic analysis of cyclosporin a binding to cyclophilin a in a lung tumor tissue lysate.

Journal Article Anal Chem · August 1, 2004 We report on the application of SUPREX (stability of unpurified proteins from rates of H/D exchange) to the analysis of a protein-ligand binding interaction under the ex vivo solution conditions of a human lung tumor tissue lysate. A SUPREX-derived binding ... Full text Link to item Cite

Facile chemical synthesis and equilibrium unfolding properties of CopG.

Journal Article Protein Sci · July 2004 The 45-amino acid polypeptide chain of the homodimeric transcriptional repressor, CopG, was chemically synthesized by stepwise solid phase peptide synthesis (SPPS) using a protocol based on Boc-chemistry. The product obtained from the synthesis was readily ... Full text Link to item Cite

The protein backbone makes important contributions to 4-oxalocrotonate tautomerase enzyme catalysis: understanding from theory and experiment.

Journal Article Biochemistry · June 2004 The role of polypeptide backbone interactions in 4-oxalocrotonate tautomerase (4OT) catalysis has been investigated using a combination of site-directed mutagenesis experiments with unnatural amino acids and quantum mechanical/molecular mechanical (QM/MM) ... Full text Cite

High-throughput screening assay for the tunable selection of protein ligands.

Journal Article Journal of combinatorial chemistry · March 2004 Here, we describe a new protein-ligand binding assay that is amenable to high-throughput screening applications. The assay involves the use of SUPREX (stability of unpurified proteins from rates of H/D exchange), a new H/D exchange and mass spectrometry-ba ... Full text Cite

A new H/D exchange- and mass spectrometry-based method for thermodynamic analysis of protein-DNA interactions.

Journal Article Chemistry & biology · December 2003 The application of SUPREX (stability of unpurified proteins from rates of H/D exchange) to the thermodynamic analysis of protein-DNA complexes is described. A series of five model protein-DNA complexes involving two known DNA binding proteins, Arc represso ... Full text Cite

Identification and validation of a potential lung cancer serum biomarker detected by matrix-assisted laser desorption/ionization-time of flight spectra analysis.

Journal Article Proteomics · September 2003 Many abnormalities detected in the thorax by routine conventional imaging studies are benign, yet all require further evaluation because of the concern for cancer. To address this deficiency and develop a serum biomarker for lung cancer, we designed a matr ... Full text Link to item Cite

Analysis of human serum proteins by liquid phase isoelectric focusing and matrix-assisted laser desorption/ionization-mass spectrometry.

Journal Article Proteomics · September 2003 Direct matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis of human serum yielded ion signals from only a fraction of the total number of peptides and proteins expected to be in the sample. We increased the ... Full text Link to item Cite

Exploring the proteome with MALDI-TOF

Journal Article Proteomics · September 1, 2003 Cite

A combinatorial approach in designing hydrophilic surfaces for solid-phase peptide synthesis

Journal Article Journal of Applied Polymer Science · July 1, 2003 A hydrophilic surface suitable for solid-state peptide synthesis was developed on a solid support called a lantern. The split-and-mix combinatorial technique was used to prepare about 500 surfaces in a very short time. Surfaces were analyzed according to v ... Full text Cite

Comparative analysis of two different amide-to-ester bond mutations in the beta-sheet of 4-oxalocrotonate tautomerase.

Journal Article Biochemistry · June 2003 Here we describe the total chemical synthesis and biophysical characterization of two backbone-modified, ester bond-containing analogues of the homohexameric enzyme 4-oxalocrotonate tautomerase (4OT). The amide-to-ester bond mutations in the two analogues ... Full text Cite

Accuracy and precision of a new H/D exchange- and mass spectrometry-based technique for measuring the thermodynamic properties of protein-peptide complexes.

Journal Article Biochemistry · May 2003 A new H/D exchange- and MALDI mass spectrometry-based technique, termed SUPREX, was used to characterize the thermodynamic properties of a series of model protein-peptide complexes of the Abelson tyrosine kinase SH3 domain (abl-SH3) and the S-Protein (S-Pr ... Full text Cite

Protein expression profiling identifies macrophage migration inhibitory factor and cyclophilin a as potential molecular targets in non-small cell lung cancer.

Journal Article Cancer Res · April 1, 2003 Current diagnostic and therapeutic strategies for lung cancer have had no significant impact on lung cancer mortality over the last several decades. This study used a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF M ... Link to item Cite

Synthesis of tetrahydro-1,4-benzodiazepine-2-ones on hydrophilic polyamide SynPhase lanterns.

Journal Article Journal of combinatorial chemistry · March 2003 Solid-phase synthesis is greatly dependent on the solid support. Here, we report the use of a new hydrophilic grafted surface on SynPhase lanterns in solid-phase organic chemistry. A convenient and facile solid-phase synthesis of disubstituted 1,4-benzodia ... Full text Cite

Use of mixture models in MALDI-TOF proteomic data for peak registration

Conference PROCEEDINGS OF THE 7TH JOINT CONFERENCE ON INFORMATION SCIENCES · January 1, 2003 Link to item Cite

A fractionation protocol to reduce signal suppression effects in the MALDI analysis of blood proteins

Journal Article Proceedings 50th Asms Conference on Mass Spectrometry and Allied Topics · December 1, 2002 A protocol involving the combination of liquid-phase isoelectric focusing and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was developed. Liquid-phase isoelectric focusing was used to separate serum peptides a ... Cite

A new H/D exchange- and MALDI-based assay for the quantitative analysis of protein-DNA interactions in solution

Journal Article Proceedings 50th Asms Conference on Mass Spectrometry and Allied Topics · December 1, 2002 A new H/D exchange- and MALDI-based assay for the quantitative analysis of protein-DNA interactions in solution was described. The protein and protein-DNA complexes were diluted ten-fold into a series of deuterated exchange buffers differing only in the co ... Cite

Quantitative protein stability measurement in living bacteria

Journal Article Proceedings 50th Asms Conference on Mass Spectrometry and Allied Topics · December 1, 2002 Hydrogen exchange detected by MALDI mass spectrometry was used to measure the thermodynamic stability of proteins. The stability of the N-terminal domain measured in vitro, qualitatively correlated with its degradation because the folding kinetics of the p ... Cite

Quantitative determination of protein-peptide binding constants using a new, MALDI- and H/D-exchange based technique

Journal Article Proceedings 50th Asms Conference on Mass Spectrometry and Allied Topics · December 1, 2002 A new MALDI-based technique, stability of unpurified proteins by rates of H/D exchange (SUPREX), for quantitative determination of protein-peptide binding constants was discussed. For the study the five different type of peptide sequence were used, which i ... Cite

A general mass spectrometry-based method for the quantitation of protein-ligand binding interactions in solution

Journal Article Proceedings 50th Asms Conference on Mass Spectrometry and Allied Topics · December 1, 2002 The protein-ligand binding interactions in solution were investigated using mass spectrometry. The H/D exchange and mass spectrometry-based technique termed as stability of unpurified proteins by rates of H/D exchange (SUPREX) was developed to evaluate the ... Cite

A general mass spectrometry-based assay for the quantitation of protein-ligand binding interactions in solution.

Journal Article J Am Chem Soc · September 4, 2002 A new method that utilizes matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and exploits the hydrogen/deuterium (H/D) exchange properties of proteins was developed for measuring the thermodynamic properties of protein-ligand complexes ... Full text Link to item Cite

Thermodynamic stability measurements on multimeric proteins using a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based method.

Journal Article Protein science : a publication of the Protein Society · April 2002 We recently reported on a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based technique, termed SUPREX, that removes several important limitations associated with measuring the thermodynamic stability of protei ... Full text Cite

A stable dimer in the pH-induced equilibrium unfolding of the homo-hexameric enzyme 4-oxalocrotonate tautomerase (4-OT).

Journal Article Biochemistry · April 2002 4-Oxalocrotonate tautomerase (4-OT) is a multimeric, bacterial enzyme comprised of 6 identical 62-amino acid subunits, which associate under native conditions to form a homo-hexameric structure stabilized entirely by noncovalent interactions. We have previ ... Full text Cite

Measurements of protein stability by H/D exchange and matrix-assisted laser desorption/ionization mass spectrometry using picomoles of material.

Journal Article Analytical chemistry · July 2001 Recently, we reported on a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI)-based technique, termed SUPREX, that can be used to measure the thermodynamic stability of a protein (Ghaemmaghami, S.; Fitzgerald, M. C.; Oas, T. G. Proc. ... Full text Cite

Guanidine-induced equilibrium unfolding of a homo-hexameric enzyme 4-oxalocrotonate tautomerase (4-OT).

Journal Article Biochemistry · April 10, 2001 4-Oxalocrotonate tautomerase (4-OT) is a bacterial enzyme that is comprised of 6 identical 62 amino acid subunits. The 4-OT enzyme is an attractive model system in which to study the interrelationship between protein folding, subunit assembly, and catalyti ... Full text Link to item Cite

A solid sample preparation method that reduces signal suppression effects in the MALDI analysis of peptides.

Journal Article Analytical chemistry · February 2001 Here we report on the application of a solid-solid (SS) sample preparation protocol for the MALDI analysis of peptides and multicomponent peptide mixtures. Our results with a series of model peptides indicate that a SS MALDI sample preparation protocol is ... Full text Cite

Identification of an essential backbone amide bond in the folding and stability of a multimeric enzyme

Journal Article Journal of the American Chemical Society · August 30, 2000 Here we utilize a total chemical synthesis strategy and a mass spectrometry-based, combinatorial chemistry approach to identify key molecular interactions that contribute to the folding and stability of a model multimeric enzyme, 4-oxalocrotonate tautomera ... Cite

Identification of an essential backbone amide bond in the folding and stability of a multimeric enzyme

Journal Article Journal of the American Chemical Society · August 30, 2000 Here we utilize a total chemical synthesis strategy and a mass spectrometry-based, combinatorial chemistry approach to identify key molecular interactions that contribute to the folding and stability of a model multimeric enzyme, 4-oxalocrotonate tautomera ... Full text Cite

A quantitative, high-throughput screen for protein stability.

Journal Article Proc Natl Acad Sci U S A · July 18, 2000 In proteomic research, it is often necessary to screen a large number of polypeptides for the presence of stable structure. Described here is a technique (referred to as SUPREX, stability of unpurified proteins from rates of H/D exchange) for measuring the ... Full text Link to item Cite

Identification of an essential backbone amide bond in the folding and stability of a multimeric enzyme

Journal Article Journal of the American Chemical Society · 2000 Here we utilize a total chemical synthesis strategy and a mass spectrometry-based, combinatorial chemistry approach to identify key molecular interactions that contribute to the folding and stability of a model multimeric enzyme, 4-oxalocrotonate tautomera ... Cite

Characterization of the role of the amino-terminal proline in the enzymatic activity catalyzed by macrophage migration inhibitory factor.

Journal Article Biochemistry · July 1998 The cytokine macrophage migration inhibitory factor (MIF) mediates several immune and inflammatory processes through unknown or poorly understood mechanisms. The protein shares structural homology with two bacterial isomerases, 4-oxalocrotonate tautomerase ... Full text Cite

Synthesis, Characterization, and Photochemical/Photophysical Properties of Ruthenium(II) Complexes with Hexadentate Bipyridine and Phenanthroline Ligands

Journal Article Inorganic Chemistry · January 1, 1998 Three hexadentate, podand-type, polypyridyl ligands, (5-bpy-2C)3Bz, (4-bpy-2C-Ph)3Et, and (4-phen-2C-Ph)3Et, and their Ru(II) and Fe(II) complexes have been prepared. Reaction of these ligands with Fe(II) produces only the ... Full text Cite

Inactivation of 4-oxalocrotonate tautomerase by 2-oxo-3-pentynoate.

Journal Article Biochemistry · December 1997 The compound, 2-oxo-3-pentynoate, has been synthesized and tested as an inhibitor of the enzyme 4-oxalocrotonate tautomerase. The enzyme is rapidly and irreversibly inactivated by the acetylenic product analogue in a time-dependent fashion. The enzyme disp ... Full text Cite

A continuous fluorometric assay for the feline immunodeficiency virus protease.

Journal Article Analytical biochemistry · December 1997 A novel fluorogenic substrate for continuous feline immunodeficiency virus (FIV) protease (PR) assay was developed in which 2-aminobenzoic acid (Abz) and p-nitrophenylalanine (F(NO2)) were used as the fluorescent donor and acceptor, respectively. The 14-am ... Full text Cite

Direct monitoring of organic reactions on polymeric supports

Journal Article Tetrahedron Letters · September 8, 1997 A method to use matrix-assisted laser desorption/ionization mass spectrometry MALDI-MS) for real time monitoring of organic reactions on polymeric supports used in solid-phase synthesis is described. The strategy utilizes a synthetic construct that allows ... Full text Cite

Methods for the chemical synthesis and readout of self-encoded arrays of polypeptide analogues

Journal Article Journal of the American Chemical Society · August 27, 1997 The synthesis of defined arrays of polypeptide analogues in conjunction with a simple self-encoded chemical readout system provides a powerful method for the systematic investigation of the relationship between polypeptide molecular structure and function. ... Full text Cite

Molecular analysis of the feline immunodeficiency virus protease: generation of a novel form of the protease by autoproteolysis and construction of cleavage-resistant proteases.

Journal Article Journal of virology · July 1997 The feline immunodeficiency virus (FIV) protease is essential for virion maturation and subsequent viral replication in that it cleaves the Gag and Gag/Pol polyproteins at eight sites to release the respective structural proteins and enzymes. During purifi ... Full text Cite

Mass spectrometry as a readout of protein structure and function.

Journal Article Mass spectrometry reviews · July 1997 Proteins have evolved to carry out very specific functions within the cell by interacting with a diverse set of biomolecules. Understanding how a protein's higher order structure relates to its function is important for defining the molecular basis of thes ... Full text Cite

A Continuous Fluorometric Assay for the FIV Protease

Journal Article Analytical Biochemistry · 1997 Cite

C2-symmetrical tetrahydroxyazepanes as inhibitors of glycosidases and HIV/FIV proteases.

Journal Article Bioorganic & medicinal chemistry · December 1996 C2-Symmetrical tetrahydroxyazepanes were synthesized as inhibitors for glycosidases. Tetrahydroxyazepane 1 is a non-specific inhibitor of various glycosidases, while compounds 2, 3 and 4 specifically inhibit beta-N-acetylglucosaminidase, beta-glucosidase, ... Full text Cite

Probing the chemical basis of binding activity in an SH3 domain by protein signature analysis.

Journal Article Chemistry & biology · October 1996 BackgroundModifying the covalent structure of a protein is an effective empirical route to probing three-dimensional structure and biological function. Here we describe a combinatorial protein chemistry strategy for studying structure-activity rel ... Full text Cite

Biochemical mass spectrometry: worth the weight?

Journal Article Chemistry & biology · September 1996 The utility of mass spectrometry for the analysis of biological molecules has been enhanced by the development of two techniques that generate gas-phase ions via nondestructive vaporization and ionization. These techniques can be used not only to determine ... Full text Cite

Probing the oligomeric structure of an enzyme by electrospray ionization time-of-flight mass spectrometry.

Journal Article Proceedings of the National Academy of Sciences of the United States of America · July 1996 Electrospray ionization time-of-flight (ESI-TOF) mass spectrometry was used to study the quaternary structure of 4-oxalocrotonate tautomerase (EC 5.3.2; 4OT), and four analogues prepared by total chemical synthesis. Wild-type 4OT is a hexamer of 62 amino a ... Full text Cite

Direct characterization of solid phase resin-bound molecules by mass spectrometry

Journal Article Bioorganic and Medicinal Chemistry Letters · April 23, 1996 In this work we demonstrate that analytes, covalently linked to a polymeric support through a photolabile linker, can be directly analyzed by matrix-assisted laser desorption/ionization (MALDI). Mass spectral analysis is performed in a single step requirin ... Full text Cite

Oligodeoxynucleotide Fragmentation in MALDI/TOF Mass Spectrometry Using 355-nm Radiation

Journal Article Journal of the American Chemical Society · January 1, 1995 The fragmentation of small oligodeoxynucleotides using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry with 355-nm radiation from the matrix 2, 5-dihydroxybenzoic acid is studied. Negative ion mass spectra of the homopolymer oligodeox ... Full text Cite

Total Chemical Synthesis and Catalytic Properties of the Enzyme Enantiomers L- and D-4-Oxalocrotonate Tautomerase

Journal Article Journal of the American Chemical Society · January 1, 1995 Both the native L-form and the mirror image D-form of the enzyme 4-oxalocrotonate tautomerase (40T) were prepared by total chemical synthesis. Our results indicate that both enzymes were efficient catalysts and demonstrate, as expected, that the achiral su ... Full text Cite

Mass spectrometry of nucleic acids: the promise of matrix-assisted laser desorption-ionization (MALDI) mass spectrometry.

Journal Article Annual review of biophysics and biomolecular structure · January 1995 In the past several years, significant progress has been made in the application of matrix-assisted laser desorption-ionization (MALDI) mass spectrometry to the analysis of large biopolymers, including nucleic acids. By isolating analyte molecules in an ap ... Full text Cite

Effect of acid predissolution on fibril size and fibril flexibility of synthetic beta-amyloid peptide.

Journal Article Biophysical journal · September 1994 beta-amyloid peptide (A beta) is the major protein component of senile plaques and cerebrovascular amyloid deposits in Alzheimer's patients. Several researchers have demonstrated that A beta is neurotoxic in in vitro and in vivo systems. Peptide aggregatio ... Full text Cite

Basic matrices for the matrix-assisted laser desorption/ionization mass spectrometry of proteins and oligonucleotides.

Journal Article Analytical chemistry · November 1993 In order to examine the importance of pH in the matrix-assisted laser desorption/ionization (MALDI) analysis of proteins and oligonucleotides, 37 highly substituted pyrimidine, pyridine, and benzene derivatives containing basic amino groups were screened a ... Full text Cite

Rapid shotgun cloning utilizing the two base recognition endonuclease CviJI.

Journal Article Nucleic acids research · July 1992 A new approach has been developed for the rapid fragmentation and fractionation of DNA into a size suitable for shotgun cloning and sequencing. The restriction endonuclease CviJI normally cleaves the recognition sequence PuGCPy between the G and C to leave ... Full text Cite