Functional Purification and Enzymic Characterization of the RNA-dependent DNA Polymerase of Human Immunodeficiency Virus
1986; Microbiology Society; Volume: 67; Issue: 12 Linguagem: Inglês
10.1099/0022-1317-67-12-2791
ISSN1465-2099
AutoresEwald M. Wondrak, Johannes Löwer, Reinhard Kurth,
Tópico(s)HIV/AIDS drug development and treatment
ResumoSummary The RNA-dependent DNA polymerase (RDDP) of human immunodeficiency virus (HIV) was purified from sucrose density gradient-banded virus by four successive procedures: anion exchange chromatography, cation exchange chromatography, affinity chromatography on oligo(dT)-cellulose and adsorption chromatography on hydroxyapatite. The enzyme preparation was free of cellular DNA-dependent DNA polymerase activity. The properties of HIV RDDP were determined with a variety of template-primers. Generally, the enzyme used Mg2+ for optimal activity except with (Cm)n.(dG)12–18 as template-primer. Kinetic data (Michaelis constant, Hill coefficient) were calculated for several substrates.
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