Luminescent and substrate binding activities of firefly luciferase N-terminal domain.
Luminescent and substrate binding activities of firefly luciferase N-terminal domain.
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萤火虫荧光素酶 N 末端结构域的发光和底物结合活性。
DOI：
10.1016/s1570-9639(03)00179-1
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Teruyuki Nagamune
中科院分区：
文献类型：
--
作者：
T. Zako;Keiichi Ayabe;T. Aburatani;N. Kamiya;A. Kitayama;H. Ueda;Teruyuki Nagamune
萤火虫荧光素酶催化从底物荧光素、Mg-ATP和氧高效发光。许多氨基酸残基被鉴定为对于发光活性是重要的，并且几乎所有的关键残基被认为位于N-末端结构域（1-437）中，除了C-末端结构域中的一个，Lys 529，其被认为对于有效的底物定向是关键的。在这里，我们表明，纯化的N-末端结构域仍然结合到底物的ATP和ATP的亲和力降低，并保留高达0.03%的野生型酶（WT）的发光活性，这表明所有的必需残基的活性位于N-末端结构域。还发现辅酶A（CoA）的低发光增强，这意味着比WT酶中更低的产物抑制。这些发现对该酶的发光反应机制，如反应中间体，产物抑制，以及C-末端结构域的作用具有有趣的意义。
Firefly luciferase catalyzes highly efficient emission of light from the substrates luciferin, Mg-ATP, and oxygen. A number of amino acid residues are identified to be important for the luminescent activity, and almost all the key residues are thought to be located in the N-terminal domain (1–437), except one in the C-terminal domain, Lys529, which is thought to be critical for efficient substrate orientation. Here we show that the purified N-terminal domain still binds to the substrates luciferin and ATP with reduced affinity, and retains luminescent activity of up to 0.03% of the wild-type enzyme (WT), indicating that all the essential residues for the activity are located in the N-terminal domain. Also found is low luminescence enhancement by coenzyme A (CoA), which implies a lower product inhibition than in the WT enzyme. These findings have interesting implications for the light emission reaction mechanism of the enzyme, such as reaction intermediates, product inhibition, and the role of the C-terminal domain.
影响因子：
2.9
作者：
PAZZAGLI, M;DEVINE, JH;BALDWIN, TO
通讯作者：
BALDWIN, TO