[HTML][HTML] Substrate requirements of human rhinovirus 3C protease for peptide cleavage in vitro.
MG Cordingley, PL Callahan, VV Sardana… - Journal of Biological …, 1990 - Elsevier
MG Cordingley, PL Callahan, VV Sardana, VM Garsky, RJ Colonno
Journal of Biological Chemistry, 1990•ElsevierA series of synthetic peptides representing authentic proteolytic cleavage sites of human
rhinovirus type 14 were assayed as substrates for purified 3C protease. Competition
cleavage assays were employed to determine the relative specificity constants (Kcat/Km) for
substrates with sequences related to the viral 2C-3A cleavage site. Variable length peptides
representing the 2C-3A cleavage site were cleaved with comparable efficiency. These
studies defined a minimum substrate of 6 amino acids (TLFQ/GP), although retention of the …
rhinovirus type 14 were assayed as substrates for purified 3C protease. Competition
cleavage assays were employed to determine the relative specificity constants (Kcat/Km) for
substrates with sequences related to the viral 2C-3A cleavage site. Variable length peptides
representing the 2C-3A cleavage site were cleaved with comparable efficiency. These
studies defined a minimum substrate of 6 amino acids (TLFQ/GP), although retention of the …
A series of synthetic peptides representing authentic proteolytic cleavage sites of human rhinovirus type 14 were assayed as substrates for purified 3C protease. Competition cleavage assays were employed to determine the relative specificity constants (Kcat/Km) for substrates with sequences related to the viral 2C-3A cleavage site. Variable length peptides representing the 2C-3A cleavage site were cleaved with comparable efficiency. These studies defined a minimum substrate of 6 amino acids (TLFQ/GP), although retention of the residue at position P5 (ETLFQ/GP) resulted in a better substrate by an order of magnitude. Amino acid substitutions at position P5, P4, P1', or P2' indicated that the identity of the residue at position P5 was not critical, whereas substitutions at position P4, P1' or P2' resulted in substrates with Kcat/Km values varying over 2 orders of magnitude. In contrast to the 2C-3A cleavage site, small peptide derivatives representative of the 3A-3B cleavage site were relatively poor substrates, which suggested that residues flanking the minimum core sequence may influence susceptibility to cleavage. The 3C protease of rhinovirus type 14 was also capable of cleaving peptides representing comparable cleavage sites predicted for coxsackie B virus and poliovirus.
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