Le Grice S F, Mills J, Mous J
Central Research Units, F. Hoffmann-La Roche & Co. Ltd., Basel, Switzerland.
EMBO J. 1988 Aug;7(8):2547-53. doi: 10.1002/j.1460-2075.1988.tb03103.x.
Replacement of the putative active site Asp residue of cloned HIV-1 protease with Ala yields a molecule incapable of autocatalytic processing. Similarly, protease/reverse transcriptase and protease/reverse transcriptase/endonuclease polyproteins containing the same mutation accumulate as enzymatically inert polyproteins. Introduction of a second, wild-type, copy of protease in trans alleviates this defect, leading in the case of individually cloned protease to cleavage of the mutant protein, and with the polyprotein mutants to release of the reverse transcriptase and endonuclease polypeptides, the former of which recover enzymatic activity. In related experiments, a similar inhibition and trans-complementation of a genetically engineered gag--protease fusion protein was observed.
将克隆的HIV-1蛋白酶假定的活性位点天冬氨酸残基替换为丙氨酸，会产生一个无法进行自身催化加工的分子。同样，含有相同突变的蛋白酶/逆转录酶和蛋白酶/逆转录酶/核酸内切酶多聚蛋白会作为无酶活性的多聚蛋白积累。反式引入第二个野生型蛋白酶拷贝可缓解此缺陷，对于单独克隆的蛋白酶而言，会导致突变蛋白的切割，对于多聚蛋白突变体而言，则会释放出逆转录酶和核酸内切酶多肽，其中前者恢复酶活性。在相关实验中，观察到了对基因工程改造的gag-蛋白酶融合蛋白的类似抑制作用和顺式互补作用。
