PURIFICATION OF HIV-1 WILD-TYPE PROTEASE AND CHARACTERIZATION OF PROTEOLYTICALLY INACTIVE HIV-1 PROTEASE MUTANTS BY PEPSTATIN-A AFFINITY-CHROMATOGRAPHY

被引：15

作者：

WONDRAK, EM ^{[1
]}

LOUIS, JM ^{[1
]}

MORA, PT ^{[1
]}

OROSZLAN, S ^{[1
]}

机构：

[1] NCI, DIV CANC BIOL & DIAG, BETHESDA, MD 20892 USA

来源：

FEBS LETTERS | 1991年 / 280卷 / 02期

关键词：

HIV-1; PROTEASE; MUTANT PROTEASE; PEPSTATIN-A AFFINITY CHROMATOGRAPHY;

D O I：

10.1016/0014-5793(91)80328-Z

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Recombinant wild-type protease of human immunodeficiency virus, type 1 (HIV-1) expressed in E. coli was purified by pepstatin A affinity chromatography. An 88-fold purification was achieved giving a protease preparation with a specific enzymatic activity of approximately 3700 pmol/min/mu-g. Two proteolytically inactive HIV-1 mutant proteases (Arg-87 --> Lys; Asn-88 --> Glu) were found to bind to pepstatin A agarose, and they were purified as the wild-type protease. A third mutant protease (Arg-87 --> Glu) was apparently unable to bind to pepstatin A under similar conditions. Binding to pepstatin A indicates the binding ability of the substrate binding site and the ability to form dimers. These features may be used to purify and to characterize other mutated HIV-1 proteases.

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页码：347 / 350

页数：4

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