Roles of amino acid residues H66 and D326 in the creatine kinase activity and structural stability.
Roles of amino acid residues H66 and D326 in the creatine kinase activity and structural stability.
复制标题
氨基酸残基 H66 和 D326 在肌酸激酶活性和结构稳定性中的作用。
DOI：
10.1016/j.ijbiomac.2017.09.020
复制
发表时间：
2018-02
影响因子：
8.2
通讯作者：
Xu Kai-Lin
中科院分区：
文献类型：
--
作者：
Wu Qing-Yun;Wei Fang;Zhu Yuan-Yuan;Tong Yu-Xue;Cao Jiang;Zhou Ping;Li Zhen-Yu;Zeng Ling-Yu;Li Feng;Wang Xiao-Yun;Xu Kai-Lin
肌酸激酶（CK）是脊椎动物细胞能量代谢的关键酶，催化磷酸肌酸向ADP的可逆磷酰转移。CK含有一对高度保守的氨基酸（H66和D326），其可能通过连接其N-和C-末端结构域而在维持CK的紧凑结构中起重要作用，但其机制尚不清楚。本研究通过光谱、结构建模和蛋白质折叠实验表明，D326 A、H66 P和H66 P/D326 A突变导致这两个氨基酸残基之间的氢键断裂，形成部分未折叠状态，在环境胁迫下更容易失活和未折叠，更容易形成不溶性聚集体。不溶性聚集体的形成会降低活性CK的水平，这可能为CK缺乏症提供线索。这些结果表明，协同作用的程度与CK的构象变化密切相关。因此，我们的研究结果提供了线索，了解氨基酸残基以外的活性位点在调节底物协同作用的机制。
Creatine kinase (CK) is a key enzyme for cellular energy metabolism, catalyzing the reversible phosphoryl transfer from phosphocreatine to ADP in vertebrates. CK contains a pair of highly conserved amino acids (H66 and D326) which might play an important role in sustaining the compact structure of CK by linking its N- and C- terminal domains; however the mechanism is still unclear. In this study, spectroscopic, structural modeling and protein folding experiments suggested that D326A, H66P and H66P/D326A mutations led to disruption of the hydrogen bond between those two amino acid residues and form the partially unfolded state which made it easier to be inactivated and unfolded under environmental stresses, and more prone to form insoluble aggregates. The formation of insoluble aggregates would decrease levels of active CKs which may provide clues in CK deficiency disease. Moreover, these results indicated that the degree of synergism had closely relationship to the conformational changes of CK. Thus, our results provided clues for understanding the mechanism of amino acid residues outside the active site in regulating substrate synergism.
登录
查看更多内容
N-和C-末端结构域之间的连接子在兔肌肉肌酸激酶的稳定性和折叠中的作用
DOI：
10.1016/j.biocel.2007.04.028
发表时间：
2007-01-01
影响因子：
4
作者：
He, Hua-Wei;Feng, Shan;Yan, Yong-Bin
通讯作者：
Yan, Yong-Bin
影响因子：
2.9
作者：
W. R. Novak;Pan‐Fen Wang;M. McLeish;G. L. Kenyon;P. Babbitt
通讯作者：
W. R. Novak;Pan‐Fen Wang;M. McLeish;G. L. Kenyon;P. Babbitt
影响因子：
3.9
作者：
Qing-yun Wu;Feng Li;Xiao-yun Wang;Zheng Chen
通讯作者：
Qing-yun Wu;Feng Li;Xiao-yun Wang;Zheng Chen
影响因子：
3.7
作者：
Povarova OI;Kuznetsova IM;Turoverov KK
通讯作者：
Turoverov KK
影响因子：
9.2
作者：
T. Wallimann
通讯作者：
T. Wallimann