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Enrichment of peptides containing consensus sequence by an enzymatic approach for targeted analysis ofproteins
Author(s) -
Wang Chunli,
Ye Mingliang,
Han Guanghui,
Chen Rui,
Zhang Manyu,
Jiang Xinning,
Cheng Kai,
Wang Fangjun,
Zou Hanfa
Publication year - 2011
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/pmic.201100041
Subject(s) - consensus sequence , sequence motif , peptide sequence , biochemistry , peptide , sequence analysis , biology , computational biology , protein sequencing , enzyme , sequence (biology) , multiple sequence alignment , chemistry , sequence alignment , gene
Multiple residues with consensus sequence, i.e. motif, on proteins are closely related to protein function. However, there is no effective method for targeted analysis of such proteins. The challenge for analysis of these classes of proteins by MS is how to selectively enrich peptides containing consensus sequence from protein digest. Although enrichment of peptides containing one type of amino acid residue was successfully achieved by chemically labeling followed by chromatographic isolation, however, it is almost impossible to label and isolate signature peptides containing multiple residues with consensus sequence by chemical approach. Herein, we developed an enzymatic approach based on the specific recognition between enzyme and its substrates to enrich such peptides. This approach was realized by modification of a residue in the consensus sequence via enzyme that can recognize the sequence followed by the isolation of the modified peptides. cAMP‐dependent protein kinase was used to validate this approach and 168 peptides containing consensus motif were identified with selectivity of 67.2%. Those peptides resulted in the identification of 88 proteins with consensus sequence from serum sample. As this motif‐oriented peptide enrichment approach allows targeted analysis of a subset of proteins with consensus sequence, it will have broad application in biological studies.