A SOLID-PHASE ASSAY FOR THE PROTEASE OF HUMAN-IMMUNODEFICIENCY-VIRUS

被引：19

作者：

WONDRAK, EM ^{[1
]}

COPELAND, TD ^{[1
]}

LOUIS, JM ^{[1
]}

OROSZLAN, S ^{[1
]}

机构：

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

来源：

ANALYTICAL BIOCHEMISTRY | 1990年 / 188卷 / 01期

关键词：

D O I：

10.1016/0003-2697(90)90530-M

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A solid phase assay for human immunodeficiency virus (HIV) protease using an immobilized substrate, Affi Gel 10-Gly-Gly-Gly-Gly-Val-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-[3H]Gly-OH has been devised. The Tyr-Pro bond of the substrate was hydrolyzed by the protease, releasing the radiolabeled cleavage product, Pro-Ile-Val-Gln-[3H]Gly-OH, into the supernatant. The pH optimum was found to be 6.0, and a high ionic strength was required for maximal activity. The solid phase assay is usable for convenient monitoring of purification procedures, and rapid screening of inhibitors of HIV protease. © 1990.

引用

收藏

页码：82 / 85

页数：4

相关论文

共 17 条

[1]

BILLICH S, 1988, J BIOL CHEM, V263, P17905

[2] GENETIC-LOCUS, PRIMARY STRUCTURE, AND CHEMICAL SYNTHESIS OF HUMAN IMMUNODEFICIENCY VIRUS PROTEASE [J].

COPELAND, TD ;

OROSZLAN, S .

GENE ANALYSIS TECHNIQUES, 1988, 5 (06) :109-115

[3] A DELETION MUTATION IN THE 5' PART OF THE POL GENE OF MOLONEY MURINE LEUKEMIA-VIRUS BLOCKS PROTEOLYTIC PROCESSING OF THE GAG AND POL POLYPROTEINS [J].

CRAWFORD, S ;

GOFF, SP .

JOURNAL OF VIROLOGY, 1985, 53 (03) :899-907

[4] HIV-1 PROTEASE SPECIFICITY OF PEPTIDE CLEAVAGE IS SUFFICIENT FOR PROCESSING OF GAG AND POL POLYPROTEINS [J].

DARKE, PL ;

NUTT, RF ;

BRADY, SF ;

GARSKY, VM ;

CICCARONE, TM ;

LEU, CT ;

LUMMA, PK ;

FREIDINGER, RM ;

VEBER, DF ;

SIGAL, IS .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1988, 156 (01) :297-303

[5] PREPARATION OF MERRIFIELD-RESINS THROUGH TOTAL ESTERIFICATION WITH CESIUM SALTS [J].

GISIN, BF .

HELVETICA CHIMICA ACTA, 1973, 56 (05) :1476-1482

[6]

KATOH I, 1985, VIROLOGY, V145, P289

[7] ACTIVE HUMAN IMMUNODEFICIENCY VIRUS PROTEASE IS REQUIRED FOR VIRAL INFECTIVITY [J].

KOHL, NE ;

EMINI, EA ;

SCHLEIF, WA ;

DAVIS, LJ ;

HEIMBACH, JC ;

DIXON, RAF ;

SCOLNICK, EM ;

SIGAL, IS .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (13) :4686-4690

[8]

KRAUSSLICH HG, 1988, ANNU REV BIOCHEM, V57, P701, DOI 10.1146/annurev.bi.57.070188.003413

[9]

KRAUSSLICH HG, 1989, CURRENT COMMUNICATIO

[10] SUBSTITUTION MUTATIONS OF THE HIGHLY CONSERVED ARGININE-87 OF HIV-1 PROTEASE RESULT IN LOSS OF PROTEOLYTIC ACTIVITY [J].

LOUIS, JM ;

SMITH, CAD ;

WONDRAK, EM ;

MORA, PT ;

OROSZLAN, S .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1989, 164 (01) :30-38