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Title: Stable isotope, site-specific mass tagging for protein identification

Abstract

Proteolytic peptide mass mapping as measured by mass spectrometry provides an important method for the identification of proteins, which are usually identified by matching the measured and calculated m/z values of the proteolytic peptides. A unique identification is, however, heavily dependent upon the mass accuracy and sequence coverage of the fragment ions generated by peptide ionization. The present invention describes a method for increasing the specificity, accuracy and efficiency of the assignments of particular proteolytic peptides and consequent protein identification, by the incorporation of selected amino acid residue(s) enriched with stable isotope(s) into the protein sequence without the need for ultrahigh instrumental accuracy. Selected amino acid(s) are labeled with .sup.13C/.sup.15N/.sup.2H and incorporated into proteins in a sequence-specific manner during cell culturing. Each of these labeled amino acids carries a defined mass change encoded in its monoisotopic distribution pattern. Through their characteristic patterns, the peptides with mass tag(s) can then be readily distinguished from other peptides in mass spectra. The present method of identifying unique proteins can also be extended to protein complexes and will significantly increase data search specificity, efficiency and accuracy for protein identifications.

Inventors:
Publication Date:
Research Org.:
Los Alamos,National Security, LLC, Los Alamos, NM (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1175960
Patent Number(s):
7,125,685
Application Number:
10/985,268
Assignee:
Los Alamos,National Security, LLC (Los Alamos, NM) LANL
DOE Contract Number:
W-7405-ENG-36
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES

Citation Formats

Chen, Xian. Stable isotope, site-specific mass tagging for protein identification. United States: N. p., 2006. Web.
Chen, Xian. Stable isotope, site-specific mass tagging for protein identification. United States.
Chen, Xian. Tue . "Stable isotope, site-specific mass tagging for protein identification". United States. doi:. https://www.osti.gov/servlets/purl/1175960.
@article{osti_1175960,
title = {Stable isotope, site-specific mass tagging for protein identification},
author = {Chen, Xian},
abstractNote = {Proteolytic peptide mass mapping as measured by mass spectrometry provides an important method for the identification of proteins, which are usually identified by matching the measured and calculated m/z values of the proteolytic peptides. A unique identification is, however, heavily dependent upon the mass accuracy and sequence coverage of the fragment ions generated by peptide ionization. The present invention describes a method for increasing the specificity, accuracy and efficiency of the assignments of particular proteolytic peptides and consequent protein identification, by the incorporation of selected amino acid residue(s) enriched with stable isotope(s) into the protein sequence without the need for ultrahigh instrumental accuracy. Selected amino acid(s) are labeled with .sup.13C/.sup.15N/.sup.2H and incorporated into proteins in a sequence-specific manner during cell culturing. Each of these labeled amino acids carries a defined mass change encoded in its monoisotopic distribution pattern. Through their characteristic patterns, the peptides with mass tag(s) can then be readily distinguished from other peptides in mass spectra. The present method of identifying unique proteins can also be extended to protein complexes and will significantly increase data search specificity, efficiency and accuracy for protein identifications.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Oct 24 00:00:00 EDT 2006},
month = {Tue Oct 24 00:00:00 EDT 2006}
}

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