Mutational analysis of conserved AAA+ residues in the archaeal Lon protease from Thermoplasma acidophilum
Mutational analysis of conserved AAA+ residues in the archaeal Lon protease from Thermoplasma acidophilum
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DOI：
10.1016/j.febslet.2004.08.021
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发表时间：
2004-09-10
期刊：
影响因子：
3.5
通讯作者：
Zwickl, P
中科院分区：
文献类型：
--
作者：
Besche, H;Tamura, N;Zwickl, P
来自古细菌Thermoplasma acidophilum （TaLon）的Lon蛋白酶由一个n端atp酶（AAA(+)）结构域和一个c端Lon蛋白酶结构域组成。尽管TaLon与可溶性Lon蛋白酶的序列相关，但在其原生宿主和大肠杆菌中表达时，TaLon被证明是膜结合的。重组TaLon被纯化为具有atp酶和蛋白水解活性的功能性高分子量复合物。对保守的AAA（+）残基的突变表明，Walker A和B基序以及sensor I和sensor 2'残基是atp酶活性所必需的，而sensor 2和精氨酸手指则参与蛋白酶结构域的激活。(C) 2004年欧洲生化学会联合会。Elsevier B.V.版权所有。
The Lon protease from the archaeon Thermoplasma acidophilum (TaLon) is composed of an N-terminal ATPase associated with various cellular activities (AAA(+)) domain and a C-terminal Lon protease domain. Although related in sequence to the soluble Lon proteases, TaLon was shown to be membrane-bound in its native host and also when expressed in Escherichia coli. Recombinant TaLon was purified as a functional high-molecular weight complex displaying ATPase and proteolytic activity. Mutagenesis of conserved AAA(+) residues revealed that the Walker A and B motifs, and the sensor I and sensor 2' residues were essential for the ATPase activity, while the sensor 2 and the arginine finger were involved in activation of the protease domain. (C) 2004 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.