Mutations in the proteolytic domain of Escherichia coli protease Lon impair the ATPase activity of the enzyme
Mutations in the proteolytic domain of Escherichia coli protease Lon impair the ATPase activity of the enzyme
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大肠杆菌蛋白酶 Lon 蛋白水解结构域的突变会损害该酶的 ATP 酶活性
DOI：
10.1016/s0014-5793(98)00012-x
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发表时间：
1998
期刊：
影响因子：
3.5
通讯作者：
T. V. Rotanova
中科院分区：
文献类型：
--
作者：
Natalie N Starkova;Ekaterina P Koroleva;L. Rumsh;L. Ginodman;T. V. Rotanova
通过比较 Lon 蛋白酶的氨基酸序列，鉴定了大肠杆菌蛋白酶 Lon 的蛋白水解结构域的保守残基，该酶是经典催化三联体（H665、H667、D676 和 D743）的推定成员。通过定点诱变获得含有取代D676N、D743N、H665Y和H667Y的突变酶。突变体 D743N 保留了三磷酸腺苷 (ATP) 依赖性蛋白水解活性，从而表明 D743 不属于催化位点。同时，突变体D676N、H665Y和H667Y丧失了水解蛋白质底物的能力。这三个突变体的 ATPase 活性降低了一个数量级以上，这表明 ATPase 和蛋白水解活性位点的空间位置接近，并且它们在蛋白质降解过程中紧密相互作用。
Conserved residues of the proteolytic domain ofEscherichia coliprotease Lon, putative members of the classic catalytic triad (H665, H667, D676, and D743) were identified by comparison of amino acid sequences of Lon proteases. Mutant enzymes containing substitutions D676N, D743N, H665Y, and H667Y were obtained by site‐directed mutagenesis. The mutant D743N retained the adenosine triphosphate (ATP)‐dependent proteolytic activity, thereby indicating that D743 does not belong to the catalytic site. Simultaneously, the mutants D676N, H665Y, and H667Y lost the capacity for hydrolysis of protein substrates. The ATPase activity of these three mutants was decreased by more than an order of magnitude, which suggests a close spatial location of the ATPase and proteolytic active sites and their tight interaction in the process of protein degradation.
DOI：
10.1016/s0021-9258(17)42340-4
发表时间：
1994-01
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
L. van Dyck;D. Pearce;F. Sherman
通讯作者：
L. van Dyck;D. Pearce;F. Sherman
影响因子：
3.5
作者：
HORTON, RM;HUNT, HD;PEASE, LR
通讯作者：
PEASE, LR
来自大肠杆菌的 ATP 依赖性蛋白酶 La (lon)。
影响因子：
--
作者：
Goldberg,AL;Moerschell,RP;Chung,CH;Maurizi,MR
通讯作者：
Maurizi,MR