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Phospho‐proteomics: Evaluation of the use of enzymatic de‐phosphorylation and differential mass spectrometric peptide mass mapping for site specific phosphorylation assignment in proteins separated by gel electrophoresis
Author(s) -
Larsen Martin R.,
Sørensen Grith Lykke,
Fey Stephen J.,
Larsen Peter Mose,
Roepstorff Peter
Publication year - 2001
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(200102)1:2<223::aid-prot223>3.0.co;2-b
Subject(s) - proteomics , peptide , bottom up proteomics , phosphorylation , peptide mass fingerprinting , protein phosphorylation , mass spectrometry , chemistry , biochemistry , gel electrophoresis , quantitative proteomics , proteome , chromatography , biology , tandem mass spectrometry , protein mass spectrometry , protein kinase a , gene
Detection of phosphorylated proteins as well as assignment of the phosphorylated sites in such proteins is a major challenge in proteomics. In the present study we evaluate the use of enzymatic de‐phosphorylation in combination with differential peptide mass mapping for identification of phosphorylated peptides in peptide mixtures derived from in‐gel digested phospho‐proteins. Phospho‐peptides could be identified provided that improved sample preparation methods prior to mass spectrometric analysis were used. An attempt to identify the proteins visualized by [ 32 P] autoradiography in a proteomics study and their phosphorylation sites, demonstrated that protein identification was possible whereas reliable identification of the phospho‐peptides requires more protein than normally available in our proteomics studies.