Characterization of human immunodeficiency virus type 1 reverse transcriptase by using monoclonal antibodies: role of the C terminus in antibody reactivity and enzyme function.
We describe the production of eight monoclonal antibodies reactive with human immunodeficiency virus type 1 reverse transcriptase (RT) by immunization of mice with purified recombinant RT. These antibodies were found to react with one or the other of two regions of the enzyme and were found to be useful in immunodeficiency purification of large amounts of the enzyme. One epitope located at the C terminus of the enzyme was of particular interest, since it was present in only the larger, 66-kilodalton (kDa) RT species and not its smaller, 51-kDa counterpart. To define this epitope, a series of mutants was made which synthesized C-terminally truncated RT. These mutants indicated that the same region of the enzyme, when deleted, both removed the C-terminal epitope and drastically reduced RT activity, indicating the importance of this region in the function of the enzyme; however, even the 51-kDa enzyme component had demonstrable activity.
Tisdale
M
M
Wellcome Research Laboratories, Beckenham, Kent, United Kingdom.
Ertl
P
P
Larder
B A
BA
Purifoy
D J
DJ
Darby
G
G
Powell
K L
KL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006678:HIV / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D006825:Hybridomas; D007118:Immunoassay; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D012194:RNA-Directed DNA Polymerase / Q000032:analysis* / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011994:Recombinant Proteins; D012194:RNA-Directed DNA Polymerase