ACTIVITY OF PURIFIED BIOSYNTHETIC PROTEINASE OF HUMAN IMMUNODEFICIENCY VIRUS ON NATURAL SUBSTRATES AND SYNTHETIC PEPTIDES
ACTIVITY OF PURIFIED BIOSYNTHETIC PROTEINASE OF HUMAN IMMUNODEFICIENCY VIRUS ON NATURAL SUBSTRATES AND SYNTHETIC PEPTIDES
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DOI：
10.1073/pnas.86.3.807
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发表时间：
1989-02-01
影响因子：
11.1
通讯作者：
CARTER, CA
中科院分区：
文献类型：
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作者：
KRAUSSLICH, HG;INGRAHAM, RH;CARTER, CA
逆转录病毒衣壳蛋白和复制酶被合成为多蛋白，由病毒编码的蛋白酶蛋白水解性加工成成熟产品。我们纯化了人类免疫缺陷病毒(HIV)的蛋白酶，在大肠杆菌中表达为apprxeq。90%的纯度。纯化的酶的浓度为.apprxeq。20 nM能快速、高效、特异地切割体外合成的Gag前体蛋白。纯化的HIV蛋白酶还能诱导5种十肽底物的特异性切割，这些底物的氨基酸序列对应于HIV多蛋白中的裂解部位，而不是对应于另一种逆转录病毒的裂解部位的多肽。用不同的多肽进行的竞争实验允许对切割位点进行排序。抑制研究表明，胃抑素A对HIV蛋白水解酶有抑制作用，其IC50为0.7µm。
Retroviral capsid proteins and replication enzymes are synthesized as polyproteins that are proteolytically processed to the mature products by a virus-encoded proteinase. We have purified the proteinase of human immunodeficiency virus (HIV), expressed in Escherichia coli to .apprxeq. 90% purity. The purified enzyme at a concentration of .apprxeq. 20 nM gave rapid, efficient, and specific cleavage of an in vitro synthesized gag precursor protein. Purified HIV proteinase also induced specific cleavage of five decapeptide substrates whose amino acid sequences corresponded to cleavage sites in the HIV polyprotein but not of a peptide corresponding to a cleavage site in another retrovirus. Competition experiments with different peptides allowed a ranking of cleavage sites. Inhibition studies indicated that the HIV proteinase was inhibited by pepstatin A with an IC50 of 0.7 .mu.M.