In vitro translation to study HIV protease activity.
HIV-1 is an etiological agent of AIDS. One of the targets of the current anti-HIV-1 combination chemotherapy, called highly active antiretroviral therapy (HAART), is HIV-1 protease (PR), which is responsible for the processing of viral structural proteins and, therefore, essential for virus replication. Here, we describe an in vitro transcription/translation-based method of phenotyping HIV-1 PR. In this system, both substrate and PR for the assay can be prepared by in vitro transcription/translation. Protease activity is estimated by the cleavage of a substrate, as measured by enzyme-linked immunosorbent assay (ELISA). This assay is safe, rapid, and requires no special facility to be carried out. Our rapid phenotyping method of HIV-1 PR may help evaluate drug resistance, useful when choosing an appropriate therapeutic regiment, and could potentially facilitate the discovery of new drugs effective against HIV-1 PR.
Matsuda
Zene
Z
Research Center for Asian Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
Iga
Mutsunori
M
Miyauchi
Kosuke
K
Komano
Jun
J
Morishita
Kazuhiro
K
Okayama
Akihiko
A
Tsubouchi
Hirohito
H
D024882:Drug Resistance, Viral / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D019428:HIV Integrase Inhibitors / Q000494:pharmacology; D016333:HIV Protease / Q000096:biosynthesis* / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology*; D066298:In Vitro Techniques; D008967:Molecular Biology / Q000379:methods*; D014176:Protein Biosynthesis; D014158:Transcription, Genetic
D019428:HIV Integrase Inhibitors; D016333:HIV Protease