Weidner J R, Dunn B M
Department of Biochemistry and Molecular Biology, J. Hillis Miller Health Center, University of Florida, Gainesville 32610.
Arch Biochem Biophys. 1991 May 1;286(2):402-8. doi: 10.1016/0003-9861(91)90058-q.
Picornaviruses, such as polio, translate their entire genome as a single polyprotein which must be proteolytically processed to produce the mature viral proteins. A majority of these cleavages are catalyzed by the virus-encoded cysteine proteinase, 3C. We report here the design and synthesis of a series of oligopeptide substrates, based upon native 3C cleavage sites, for an HPLC assay of poliovirus 3C proteinase activity. A similar series of peptides based upon human rhinovirus 3C cleavage sites was also examined. The enzyme shows a marked preference for those peptides with a proline in the P'2 position. A quenched fluorescent substrate suitable for continuous assay of 3C proteinase activity was also synthesized. Both the HPLC assay and the fluorescence assay were used to evaluate a number of potential 3C proteinase inhibitors.
小核糖核酸病毒，如脊髓灰质炎病毒，将其整个基因组作为一个单一的多聚蛋白进行翻译，该多聚蛋白必须经过蛋白水解加工才能产生成熟的病毒蛋白。这些切割反应大多由病毒编码的半胱氨酸蛋白酶3C催化。我们在此报告基于天然3C切割位点设计和合成的一系列寡肽底物，用于脊髓灰质炎病毒3C蛋白酶活性的高效液相色谱（HPLC）分析。还检测了基于人鼻病毒3C切割位点的类似系列肽段。该酶对P'2位置含有脯氨酸的那些肽段表现出明显的偏好。还合成了一种适用于连续检测3C蛋白酶活性的淬灭荧光底物。HPLC分析和荧光分析都用于评估多种潜在的3C蛋白酶抑制剂。
