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PubMed · 2090647

A simple, continuous fluorometric assay for HIV protease.

M V Toth·G R Marshall
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Abstract

Novel fluorogenic substrates for human immunodeficiency viral protease have been developed based on the principle of fluorescence energy transfer. Starting from a p24/p15 cleavage site-derived hexapeptide substrate. Ac-Thr-Ile-Nle-Nle-Gln-Arg-NH2, incorporation of 2-aminobenzoic acid in place of the acetyl group as the donor and p-NO2-Phe at the P1' position as acceptor gave the intramolecularly quenched fluorogenic substrate. Cleavage of the substrate by HIV protease released the fluorescent N-terminal tripeptide from its close apposition to the quenching nitrobenzyl group, resulting in enhanced fluorescence. An automated assay based on 96-well microtiter plates and a fluorometric plate reader have been developed, which allow high throughput of compounds in the search for HIV protease inhibitors.

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Amino Acid SequenceFluorometryHIV ProteaseHumansKineticsMolecular Sequence DataSubstrate SpecificityJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.

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M V Toth, G R Marshall. 1990. A simple, continuous fluorometric assay for HIV protease.. https://doi.org/10.1111/j.1399-3011.1990.tb00994.x

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