Journal
PROTEOMICS
Volume 11, Issue 17, Pages 3578-3581Publisher
WILEY-BLACKWELL
DOI: 10.1002/pmic.201100041
Keywords
MS; Protein kinases; Proteomics methods; Selective isolation; Targeted analysis; Technology
Funding
- NSFC [21021004]
- China State Key Basic Research Program Grant [2007CB914103, 2010CB912103]
- MOST [2009IM031800, 2010IM030500]
- National Key Special Program on Infection Diseases [2008ZX10002-017, 2008ZX10002-020]
- CAS [KJCX2.YW.HO9, KSCX2.YW.079]
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available