Luminometric method for screening retroviral protease inhibitors

Dana Horáková; Michaela Rumlová; Iva Pichová; Tomás Ruml

doi:10.1016/j.ab.2005.07.013

Luminometric method for screening retroviral protease inhibitors

Anal Biochem. 2005 Oct 1;345(1):96-101. doi: 10.1016/j.ab.2005.07.013.

Authors

Dana Horáková¹, Michaela Rumlová, Iva Pichová, Tomás Ruml

Affiliation

¹ Department of Biochemistry and Microbiology, Institute of Chemical Technology, Technická 3, 166 28 Prague, Czech Republic.

PMID: 16125122
DOI: 10.1016/j.ab.2005.07.013

Abstract

We have developed a sensitive luminometric assay for determining the activity of retroviral proteases that uses proteolytic cleavage of polypeptide substrate immobilized on Ni-NTA HisSorb Strips microplates. The protease substrate derived from the Gag precursor protein of Mason-Pfizer monkey virus (M-PMV) was conjugated with horseradish peroxidase (HRP), which catalyzes oxidation of luminol in the assay. The cleavage of the substrate was monitored as a decrease in luminescent signal caused by the release of the cleavage product conjugated to HRP. Testing of a set of M-PMV protease inhibitors confirmed that this method is sufficiently sensitive and specific for high-throughput screening of retroviral protease inhibitors.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Drug Evaluation, Preclinical / methods
Endopeptidases / chemistry*
Luminescent Measurements* / methods
Mason-Pfizer monkey virus / enzymology*
Protease Inhibitors / analysis*
Substrate Specificity

Substances

Protease Inhibitors
Endopeptidases
Mason-Pfizer monkey virus protease