Skip to main content

Peptide array X-linking (PAX): a new peptide-protein identification approach.

Publication ,  Journal Article
Okada, H; Uezu, A; Soderblom, EJ; Moseley, MA; Gertler, FB; Soderling, SH
Published in: PLoS One
2012

Many protein interaction domains bind short peptides based on canonical sequence consensus motifs. Here we report the development of a peptide array-based proteomics tool to identify proteins directly interacting with ligand peptides from cell lysates. Array-formatted bait peptides containing an amino acid-derived cross-linker are photo-induced to crosslink with interacting proteins from lysates of interest. Indirect associations are removed by high stringency washes under denaturing conditions. Covalently trapped proteins are subsequently identified by LC-MS/MS and screened by cluster analysis and domain scanning. We apply this methodology to peptides with different proline-containing consensus sequences and show successful identifications from brain lysates of known and novel proteins containing polyproline motif-binding domains such as EH, EVH1, SH3, WW domains. These results suggest the capacity of arrayed peptide ligands to capture and subsequently identify proteins by mass spectrometry is relatively broad and robust. Additionally, the approach is rapid and applicable to cell or tissue fractions from any source, making the approach a flexible tool for initial protein-protein interaction discovery.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

PLoS One

DOI

EISSN

1932-6203

Publication Date

2012

Volume

7

Issue

5

Start / End Page

e37035

Location

United States

Related Subject Headings

  • Wiskott-Aldrich Syndrome Protein Family
  • Tandem Mass Spectrometry
  • Proteomics
  • Protein Interaction Domains and Motifs
  • Peptide Library
  • Nerve Tissue Proteins
  • Molecular Sequence Data
  • Mice
  • Intracellular Signaling Peptides and Proteins
  • General Science & Technology
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Okada, H., Uezu, A., Soderblom, E. J., Moseley, M. A., Gertler, F. B., & Soderling, S. H. (2012). Peptide array X-linking (PAX): a new peptide-protein identification approach. PLoS One, 7(5), e37035. https://doi.org/10.1371/journal.pone.0037035
Okada, Hirokazu, Akiyoshi Uezu, Erik J. Soderblom, M Arthur Moseley, Frank B. Gertler, and Scott H. Soderling. “Peptide array X-linking (PAX): a new peptide-protein identification approach.PLoS One 7, no. 5 (2012): e37035. https://doi.org/10.1371/journal.pone.0037035.
Okada H, Uezu A, Soderblom EJ, Moseley MA, Gertler FB, Soderling SH. Peptide array X-linking (PAX): a new peptide-protein identification approach. PLoS One. 2012;7(5):e37035.
Okada, Hirokazu, et al. “Peptide array X-linking (PAX): a new peptide-protein identification approach.PLoS One, vol. 7, no. 5, 2012, p. e37035. Pubmed, doi:10.1371/journal.pone.0037035.
Okada H, Uezu A, Soderblom EJ, Moseley MA, Gertler FB, Soderling SH. Peptide array X-linking (PAX): a new peptide-protein identification approach. PLoS One. 2012;7(5):e37035.

Published In

PLoS One

DOI

EISSN

1932-6203

Publication Date

2012

Volume

7

Issue

5

Start / End Page

e37035

Location

United States

Related Subject Headings

  • Wiskott-Aldrich Syndrome Protein Family
  • Tandem Mass Spectrometry
  • Proteomics
  • Protein Interaction Domains and Motifs
  • Peptide Library
  • Nerve Tissue Proteins
  • Molecular Sequence Data
  • Mice
  • Intracellular Signaling Peptides and Proteins
  • General Science & Technology