来自大肠杆菌的二皮里米丁脱酶:催化序列的证据
Tyler B Alt1, Aj K Croney1, Graham R Moran1
1Department of Chemistry and Biochemistry, Loyola University Chicago, 1068 W Sheridan Rd, Chicago, Illinois 60660, United States.
基胺脱酶 (DPD) 酶活性受到其FAD和FMN辅因子之间的电子转移的限制,而不是基胺的减少. 降解激活会影响酶构成和电子流.
科学领域:
- 生物化学 生物化学
- 酶学 是一种酶学.
- 分子生物学分子生物学
背景情况:
- 二皮里米丁脱酶 (DPD) 使用NADH催化皮里米丁的减少.
- 之前的研究表明,DPD经历了还原性激活,其周转率依赖于NADH.
研究的目的:
- 为了阐明DPD营业额中的限速步骤.
- 研究电子转移和酶激活在DPD功能中的作用.
主要方法:
- 线性自由能量关系分析与化 uracils.
- 使用 [4S-2H] NADH.NADH 的位点定向突变发生 (C137S) 和动态同位素效应研究.
- 光谱分析 (电荷转移吸收) 和酶失活检测.
主要成果:
- 皮里米丁减少并不是DPD营业额中的速度限制步骤.
- 从FAD到FMN的电子转移是限制速度的.
- 减少激活导致一个特定的辅因子状态 (FADH2·4(Fe4S4) ·FMN).
- 酶静电不控制还原激活;一个移动循环在激活时重新定向.
结论:
- 在FAD和FMN辅因子之间的电子传输决定了DPD催化速率.
- 酶激活涉及结构变化,包括移动循环重定向,独立于本地FAD静电.
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