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Stable isotope labelling in vivo as an aid to protein identification in peptide mass fingerprinting
Author(s) -
Pratt Julie M.,
Robertson Duncan H. L.,
Gaskell Simon J.,
RibaGarcia Isabel,
Hubbard Simon J.,
Sidhu Khushwant,
Oliver Stephen G.,
Butler Philip,
Hayes Andrew,
Petty June,
Bey Robert J.
Publication year - 2002
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/1615-9861(200202)2:2<157::aid-prot157>3.0.co;2-m
Subject(s) - peptide mass fingerprinting , proteome , stable isotope labeling by amino acids in cell culture , amino acid , bottom up proteomics , labelling , mass spectrometry , peptide , chemistry , matrix assisted laser desorption/ionization , label free quantification , isobaric labeling , chromatography , biochemistry , in vivo , tandem mass tag , isotope , protein mass spectrometry , identification (biology) , proteomics , quantitative proteomics , biology , tandem mass spectrometry , desorption , genetics , physics , botany , organic chemistry , adsorption , quantum mechanics , gene
Peptide mass fingerprinting (PMF) is a powerful technique for identification of proteins derived from in‐gel digests by virtue of their matrix‐assisted laser desorption/ionization‐time of flight mass spectra. However, there are circumstances where the under‐representation of peptides in the mass spectrum and the complexity of the source proteome mean that PMF is inadequate as an identification tool. In this paper, we show that identification is substantially enhanced by inclusion of composition data for a single amino acid. Labelling in vivo with a stable isotope labelled amino acid (in this paper, decadeuterated leucine) identifies the number of such amino acids in each digest fragment, and show a considerable gain in the ability of PMF to identify the parent protein. The method is tolerant to the extent of labelling, and as such, may be applicable to a range of single cell systems.