Kotler M, Katz R A, Danho W, Leis J, Skalka A M
Department of Molecular Oncology, Roche Institute of Molecular Biology, Nutley, NJ 07110.
Proc Natl Acad Sci U S A. 1988 Jun;85(12):4185-9. doi: 10.1073/pnas.85.12.4185.
Processing of the gag and pol gene precursor proteins of retroviruses is essential for infectivity and is directed by a viral protease that is itself included in one of these precursors. We demonstrate here that small synthetic peptides can be used as both model substrates and inhibitors to investigate the specificity and molecular parameters of the reaction. The results indicate that a peptide that extends five amino acids but not three amino acids in both directions from a known cleavage site is accurately hydrolyzed by the protease of avian sarcoma-leukosis virus. Substitutions of the amino acids to either side of the peptide bond to be cleaved affect the ability of the peptide (as well as a larger precursor protein) to serve as a substrate. The specificity is more stringent for the amino acid that will become the carboxyl end after cleavage. Some substitutions produced peptides that were not cleaved but could act as inhibitors of cleavage of a susceptible peptide. Thus, small model substrates may be used to explore both the binding and catalytic properties of these important proteases.
逆转录病毒gag和pol基因前体蛋白的加工对于感染性至关重要,且由一种病毒蛋白酶指导,而该蛋白酶本身包含在这些前体之一中。我们在此证明,小的合成肽可作为模型底物和抑制剂来研究该反应的特异性和分子参数。结果表明,从已知切割位点向两个方向延伸五个氨基酸而非三个氨基酸的肽能被禽肉瘤-白血病病毒蛋白酶精确水解。待切割肽键两侧氨基酸的取代会影响该肽(以及更大的前体蛋白)作为底物的能力。对于切割后将成为羧基末端的氨基酸,特异性更为严格。一些取代产生的肽不能被切割,但可作为敏感肽切割的抑制剂。因此,小的模型底物可用于探索这些重要蛋白酶的结合和催化特性。