Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1175649 | Analytical Biochemistry | 2009 | 5 Pages |
Abstract
The synthesis and detailed enzymatic analysis of fluorescence resonance energy transfer (FRET)-based peptides as substrates for chymopapain are reported. The design of these substrates arose from a massively parallel high-throughput microarray screening process using peptide nucleic acid (PNA) encoding technology, allowing the identification of detailed substrate specificities of any protease. Two peptides so identified with chymopapain were observed to be excellent substrates with low micromolar Km values and turnover numbers on the order of hundreds per second. Mass spectroscopy studies showed unequivocally the specificity of chymopapain toward Ala, Pro, Val, and Lys for positions P4 to P1 while not presenting high specificity for residues in position P1â².
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Juan J. Diaz-Mochon, Songsak Planonth, Mark Bradley,