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:
- Issue Date:
- Research Org.:
- Los Alamos,National Security, LLC, Los Alamos, NM (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1175960
- Patent Number(s):
- 7125685
- Application Number:
- 10/985,268
- Assignee:
- Los Alamos,National Security, LLC (Los Alamos, NM)
- Patent Classifications (CPCs):
-
G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
H - ELECTRICITY H01 - BASIC ELECTRIC ELEMENTS H01J - ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- 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. 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}
}
Works referenced in this record:
Genetic tools for selective labeling of proteins with ?-15N-amino acids
journal, September 1996
- Waugh, DavidS.
- Journal of Biomolecular NMR, Vol. 8, Issue 2
Proteome analysis using selective incorporation of isotopically labeled amino acids
journal, January 2000
- Veenstra, Timothy D.; Martinović, Suzana; Anderson, Gordon A.
- Journal of the American Society for Mass Spectrometry, Vol. 11, Issue 1