Thioltransferase (Glutaredoxin) Is Detected Within HIV-1 and Can Regulate the Activity of Glutathionylated HIV-1 Protease in Vitro

摘要: Abstract Previous studies have suggested that the two conserved cysteines of HIV-1 protease may be involved in regulating activity. Here, we examined diglutathionylated wild type (Cys-67-SSG, Cys-95-SSG) and monoglutathionylated mutants (C67A, Cys-95-SSG C95A, Cys-67-SSG) as potential substrates for thioltransferase (glutaredoxin). Time-dependent changes extent deglutathionylation each protein were assayed by reverse phase-high performance liquid chromatography. Glutathione alone was not an effective reductant, whereas displayed differential catalysis toward Cys-67-SSG sites. At low concentrations (5 nm), occurred almost exclusively at Cys-95-SSG. With substantially more (100 nm) partially deglutathionylated but only 20% rate reduction. Treatment with restored activity generated enzyme preparation had a 3- to 5-fold greater specific relative fully reduced form. Immunoblot analysis HIV-1MN virus antibody detected band co-migrating recombinant persisted following subtilisin treatment, indicating presence within HIV-1. Our results implicate regulation and/or maintenance infected cells.