A solid phase assay for the protease of human immunodeficiency virus.
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.
Wondrak
E M
EM
Laboratory of Molecular Virology and Carcinogenesis, NCI-Frederick Cancer Research Facility, Maryland 21701.
Copeland
T D
TD
Louis
J M
JM
Oroszlan
S
S
D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000201:enzymology; D006678:HIV / Q000201:enzymology*; D016333:HIV Protease / Q000032:analysis*; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000302:isolation & purification
D010455:Peptides; D016333:HIV Protease