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全长NAD+-I核糖开关结合单个辅因子,但不能歧视腺三酸盐
Yoshita Srivastava1, Maya E Blau1, Jermaine L Jenkins1
1Department of Biochemistry & Biophysics and Center for RNA Biology, University of Rochester School of Medicine & Dentistry, Rochester, New York 14642, United States.
Biochemistry
|November 10, 2023
概括
这项研究揭示了细菌I类NAD+核糖开关与单个尼古丁胺胺氨基二核酸 (NAD+) 结合,具有高亲和力. 然而, рибо开关也结合ATP,质疑其独特的NAD+特异性.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 在RNA生物学,RNA生物学.
背景情况:
- 细菌的 рибо开关通过结合小分子来调节基因表达.
- 两种类型的核糖开关感知尼古丁胺氨酸二核酸 (NAD+),这是一个关键的氧化还原辅因子.
- NAD+-I рибо开关有两个同源域,但其全长结构和功能尚不清楚.
研究的目的:
- 为了研究单个域的辅因子结合亲和性和特异性以及全长细菌NAD+-I рибо开关.
- 为了澄清这种 рибо开关类对NAD+传感的机制.
主要方法:
- 异热定位热量计 (ITC) 用于量化结合亲缘关系.
- 使用凝过色谱来评估域自我关联.
- 确定了各种二核酸的连接键结合史泰基度.
主要成果:
- NAD+ - I рибо开关的 I 域将 NAD+ 与 24.6 μM 的 KD 结合在一起,但表现出非生产性的自我关联.
- 域II显示没有可检测到与NAD+的结合.
- 全长的 рибо开关将单个NAD+与31.5μM的KD结合,并意外地将具有更高亲和力的ATP结合 (KD=11.0μM).
- 全长度 рибо开关的结合亲和顺序是ADP=ATP>NAD+=AP2-利巴维林>NADH.
结论:
- 全长的NAD+-I рибо开关将NAD+与生理相关度结合在一起.
- 观察到的ATP与其他二核酸的结合表明其特异性低于以前的假设.
- 需要进一步的研究,以了解这种双结合能力对基因调控的影响.
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