A microtiter colorimetric assay for the HIV-1 protease.
Anal Biochem
; 248(2): 246-50, 1997 Jun 01.
Article
em En
| MEDLINE
| ID: mdl-9177750
ABSTRACT
We have developed a novel colorimetric assay for the HIV-1 protease that is suitable for high-throughput screening of inhibitors. This assay utilizes two nonenzymatic reaction steps, which are carried out in succession following enzymatic hydrolysis of a synthetic peptide. The first step involves a carbamylation reaction between cyanate and the nascent alpha amino group resulting from enzymatic hydrolysis. The second step involves a carbamidodiacetyl reaction between 2,3-butanedione monoxime (diacetylmonoxime) and the de novo carbamido compound. The entire assay can be performed in a microtiter plate and is amenable to automation. In addition, this peptidolysis assay is readily adaptable to other proteolytic enzymes and their substrates.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Protease de HIV
/
Colorimetria
Idioma:
En
Revista:
Anal Biochem
Ano de publicação:
1997
Tipo de documento:
Article