Characterization of recombinant integrase of human immunodeficiency virus type 1 (isolate Bru)

Integration of the human immunodeficiency virus type 1 (HIV-1) DNA into the human genome requires the virus-encoded integrase protein. The recombinant integrase protein of HW-1 (isolate Bru) was prepared by constructing a plasmid based on pET-15b encoding the integrase gene. Integrase of HfV-1 was purified using a bacterial expression system (Escherichia coli). The main kinetic parameters of HIV-1 integrase (K-m = (3.7 +/- 0.2).10(-10) M, k(cat) = (1.2 +/- 0.3).10(-7) sec(-1)) were determined using an oligonucleotide duplex constructed on the basis of the U5-terminal sequence of proviral HIV-1 DNA as the substrate. Inhibition of integrase by aurintricarbonic acid ([I](50) = 6.3 +/- 0.4 muM) and dependence of integrase activity on Mg2+ and Mn2+ concentration were studied.