Functional purification and enzymic characterization of the RNA-dependent DNA polymerase of human immunodeficiency virus.
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 X (dG)12-18 as template-primer. Kinetic data (Michaelis constant, Hill coefficient) were calculated for several substrates.
D002845:Chromatography; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004247:DNA; D006678:HIV / Q000201:enzymology*; D006882:Hydroxyapatites; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism; D013698:Templates, Genetic
D006882:Hydroxyapatites; D004247:DNA; D012194:RNA-Directed DNA Polymerase; D008274:Magnesium