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Updated: Jul 12, 2025

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了解ATP与DOSS催化域的结合,具有短的ATP-Lid

Grant W Larson¹, Peter K Windsor¹, Elizabeth Smithwick¹

¹Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, United States.

|October 31, 2023

概括

在Mycobacterium结核病中的关键蛋白DosS的短ATP-lid不会阻止ATP结合. 这一发现挑战了之前的假设,并对理解细菌中类似蛋白质产生影响.

科学领域:

生物化学生物化学
结构生物学结构生物学
微生物学微生物学

背景情况:

DosS 是一种含有血红素的胺酶,对于*Mycobacterium tuberculosis*的休眠状态至关重要.
DosS的催化ATP结合 (CA) 域具有一个短的ATP盖,以前认为它会阻碍ATP结合.
假设这种假设性障碍需要与二元化和胺转移 (DHp) 域相互作用以进行ATP结合.

研究的目的:

重新评估DosS CA域的ATP结合机制.
为了研究短ATP-lid在核酸结合中的作用.
为了澄清DosS CA域的构造的结构和功能影响.

主要方法:

计算建模和结构生物学技术.
生物物理研究,包括循环二重化.
对DosS CA晶体结构,AlphaFold模型和同类蛋白质的分析.

主要成果:

结晶结构中的闭盖形状是一种由高度的Zn2+引起的人工物,这些度与Glu537.7协调.
Zn2+还会在N-box α螺旋中诱导一个人造的随机线圈,这对于ATP结合至关重要.
在没有Zn²⁺的情况下,DosS CA表现出结构灵活性,并与53±13μM的K_d结合AMP-PNP;结合不受DHp域结合的影响 (K_d=51±6μM).

结论:

在DOSS CA的短ATP-lid没有阻碍ATP结合.
在之前的研究中观察到的结构特征是高度Zn2+的产物.
这些发现与2988种具有类似ATP-lid结构的同类细菌蛋白相关.

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