The baculovirus cysteine protease has a cathepsin B-like S2-subsite specificity.

D Brömme, K Okamoto
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引用次数: 33

Abstract

Autographa californica nuclear polyhedrosis virus (AcNPV) encodes a functional cysteine protease of the papain family which is expressed after infection in Spodoptera fruglperda Sf9 cells. The protease displays an inhibition profile typical for cysteine proteases and is highly active against synthetic peptide substrates. The pH optimum of the bell-shaped pH-activity curve is between 5.0 and 5.5. The best substrate tested is Z-Arg-Arg-MCA which is specific for cathepsin B. The specificity constant (Kcat/Km) of AcNPV protease for this substrate is approximately two times higher than for human cathepsin B. In contrast to human cathepsins, AcNPV protease does not exhibit a discriminating specificity towards neutral hydrophobic residues in the P2 position. These substrates are hydrolysed at a ten-fold lower rate than the P2 arginine containing substrate. The pH activity profile against the Z-Arg-Arg-MCA substrate reveals a pK of 5.35 which can be assigned to a glutamate residue in the S2 subsite pocket. Like in cathepsin B, this residue facilitates the binding of positively charged P2 residues in the primary binding pocket. In this respect, the AcNPV protease resembles cathepsin B more than cathepsins L and S.

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杆状病毒半胱氨酸蛋白酶具有组织蛋白酶b样s2亚位点特异性。
加州签名虫核多角体病毒(AcNPV)编码木瓜蛋白酶家族的一种功能性半胱氨酸蛋白酶,该酶在感染后在狐尾蛾Sf9细胞中表达。该蛋白酶表现出典型的半胱氨酸蛋白酶的抑制特征,对合成肽底物具有高度活性。钟形pH-活性曲线的最适pH值为5.0 ~ 5.5。测试的最佳底物是Z-Arg-Arg-MCA,它对组织蛋白酶b具有特异性。AcNPV蛋白酶对该底物的特异性常数(Kcat/Km)约为人类组织蛋白酶b的两倍。与人类组织蛋白酶相比,AcNPV蛋白酶对P2位置的中性疏水残基没有区别性。这些底物的水解速率比含有P2精氨酸的底物低10倍。对Z-Arg-Arg-MCA底物的pH活性谱显示,pK为5.35,可归因于S2亚位口袋中的谷氨酸残基。与组织蛋白酶B一样,该残基促进了初级结合袋中带正电的P2残基的结合。在这方面,与组织蛋白酶L和S相比,AcNPV蛋白酶更像组织蛋白酶B。
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