The role of firefly luciferase C-terminal domain in efficient coupling of adenylation and oxidative steps
The role of firefly luciferase C-terminal domain in efficient coupling of adenylation and oxidative steps
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DOI：
10.1016/j.febslet.2005.07.004
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发表时间：
2005-08-15
期刊：
影响因子：
3.5
通讯作者：
Ueda, H
中科院分区：
文献类型：
--
作者：
Ayabe, K;Zako, T;Ueda, H
石楠萤火虫荧光素酶的N-末端结构域(N-结构域)具有很弱的发光活性，并呈现出独特的发光特性，其上升时间超过几分钟。通过对反应中间体荧光素腺苷(LH2-AMP)的灵敏分析，我们发现N-结构域发光的缓慢增加真实地反映了游离LH2-AMP的浓度。除了一种上升时间较长的酶外，野生型和突变型酶的上升时间较短，没有观察到这种相关性。结果表明，C-末端结构域在有效偶联腺化和氧化步骤中起着不可或缺的作用。(C)2005年欧洲生化学会联合会。爱思唯尔出版，版权所有。
The N-terminal domain (N-domain) of the firefly luciferase from Photinus pyraris has weak luminescence activity, and shows a unique light emitting profile with very long rise time of more than several minutes. Through a sensitive assay of the reaction intermediate luciferyl-adenylate (LH2-AMP), we found that the slow increase in the N-domain luminescence faithfully reflected the concentration of dissociated LH2-AMP. No such correlation was observed for wild-type or mutant enzymes with short rise time, except one with longer rise time. The results suggest that the C-terminal domain plays an indispensable role in efficiently coupling adenylation and oxidative steps. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.