Chandra P, Gerber T, Chandra A, Sarin P S, Pavlikov S P, Sidorovich I, Khaitov R, Ivanov V, Koziach A
Laboratory of Molecular Biology, University Medical School, Frankfurt am Main, FRG.
Biomed Sci. 1990;1(5):507-12.
With the aid of monoclonal antibodies to the reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1), low-molecular-mass subunits (p29, p32, and p40) were identified in HIV-1 RT purified from HIV (HTLV-IIIB) virions by isoelectric focusing. Epitope mapping with synthetic polypeptides from various regions of the pol gene suggests that the low-molecular-mass subunits result from N-terminal cleavage of the p51 subunit. The subunits could be separated only by SDS-polyacrylamide gel electrophoresis and detected by immunoblotting. They could not be separated on chromatographic columns, suggesting that the subunits are complexed or conformationally arranged in such a way that their separation on the basis of molecular mass is not possible. The molecular mass of the active enzyme eluted from a chromatographic column (Sephacryl S-300) loaded with a mixture of the subunits was estimated to be 100 kDa.
借助针对1型人类免疫缺陷病毒（HIV-1）逆转录酶（RT）的单克隆抗体，通过等电聚焦在从HIV（HTLV-IIIB）病毒粒子纯化的HIV-1 RT中鉴定出低分子量亚基（p29、p32和p40）。用来自pol基因不同区域的合成多肽进行表位作图表明，低分子量亚基是由p51亚基的N端切割产生的。这些亚基只能通过SDS-聚丙烯酰胺凝胶电泳分离，并通过免疫印迹检测。它们不能在色谱柱上分离，这表明这些亚基以一种基于分子量无法分离的方式形成复合物或构象排列。从装有亚基混合物的色谱柱（Sephacryl S-300）上洗脱的活性酶的分子量估计为100 kDa。
