Cysteine 95 and other residues influence the regulatory effects of Histidine 69 mutations on Human Immunodeficiency Virus Type 1 protease autoprocessing.
Cysteine 95 and other residues influence the regulatory effects of Histidine 69 mutations on Human Immunodeficiency Virus Type 1 protease autoprocessing.
复制标题
DOI：
10.1186/1742-4690-7-24
复制
发表时间：
2010-03-23
期刊：
影响因子：
3.3
通讯作者：
Chen C
中科院分区：
文献类型：
--
作者：
Huang L;Hall A;Chen C
HIV Gag-Pol前体的规律性自动加工是生产成熟和完全活性的蛋白酶所必需的。我们之前曾报道，在体外自动处理实验中和在转基因哺乳动物细胞中，假性野生型蛋白酶序列中的H69E突变显著阻碍了蛋白酶的成熟(>；20倍)。有趣的是，在实验室适应的NL4-3蛋白酶的背景下，H69E突变对蛋白酶成熟只显示出中等程度的抑制(~4倍)。NL4-3和假野生型蛋白水解酶之间存在6个点突变(Q7K、L33I、N37S、L63I、C67A和C95A)，表明H69E效应受到其他残基的影响。突变分析表明，C95是抑制H69E抑制作用的主要决定因素。L63和C67也表现出较小的抢救效果。然而，当NL4-3主干上的H69被天冬氨酸取代时，救援完全取消了。在H69E突变的背景下，表面残基(E21、D30、E34、E35和F99)的电荷替换为中性或带正电荷的氨基酸无法恢复蛋白酶的自动加工。综上所述，我们认为残基69与其他氨基酸如C95加L63和C67一起，在较小程度上调节了受感染细胞中蛋白水解酶自动加工的前体结构。
Regulated autoprocessing of HIV Gag-Pol precursor is required for the production of mature and fully active protease. We previously reported that H69E mutation in a pseudo wild type protease sequence significantly (>20-fold) impedes protease maturation in an in vitro autoprocessing assay and in transfected mammalian cells. Interestingly, H69E mutation in the context of a laboratory adapted NL4-3 protease showed only moderate inhibition (~4-fold) on protease maturation. There are six point mutations (Q7K, L33I, N37S, L63I, C67A, and C95A) between the NL4-3 and the pseudo wild type proteases suggesting that the H69E effect is influenced by other residues. Mutagenesis analyses identified C95 as the primary determinant that dampened the inhibitory effect of H69E. L63 and C67 also demonstrated rescue effect to a less extent. However, the rescue was completely abolished when H69 was replaced by aspartic acid in the NL4-3 backbone. Charge substitutions of surface residues (E21, D30, E34, E35, and F99) to neutral or positively charged amino acids failed to restore protease autoprocessing in the context of H69E mutation. Taken together, we suggest that residue 69 along with other amino acids such as C95 plus L63 and C67 to a less extent modulate precursor structures for the regulation of protease autoprocessing in the infected cell.
登录
查看更多内容
影响因子：
5.4
作者：
Pettit, SC;Clemente, JC;Kaplan, AH
通讯作者：
Kaplan, AH
影响因子：
64.8
作者：
LOEB, DD;SWANSTROM, R;HUTCHISON, CA
通讯作者：
HUTCHISON, CA
影响因子：
56.9
作者：
BARRESINOUSSI, F;CHERMANN, JC;MONTAGNIER, L
通讯作者：
MONTAGNIER, L
影响因子：
4.8
作者：
Koh, Yasuhiro;Matsumi, Shintaro;Mitsuya, Hiroaki
通讯作者：
Mitsuya, Hiroaki
影响因子：
5.4
作者：
Davis, DA;Yusa, K;Yarchoan, R
通讯作者：
Yarchoan, R