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结合实验和理论研究,研究长距离相互作用,调节酵母体格拉尼尔格拉尼尔二酸盐合成酶的二分化和活性

Chia-Hsiang Lo¹, Ying-Hsuan Chang, Jon D Wright

¹Institute of Biochemical Sciences, National Taiwan University, Taipei.

Journal of the American Chemical Society

|February 28, 2009

概括

基拉尼尔基拉尼尔二酸合成酶 (GGPPs) 螺旋体A中的两个氨基酸残留对酶二分化和活性至关重要. 突变破坏了二分化,显著降低了GGPPs的活动.

科学领域:

生物化学生物化学
结构生物学结构生物学
酶学是一种酶学.

背景情况:

基拉尼尔基拉尼尔二酸盐合成酶 (GGPPs) 对于合成叶绿素,胡卜素和基拉尼尔基拉尼化蛋白的前体至关重要.
GGPPs催化了法纳西二酸盐和异二酸盐的凝结,形成一个C(20) 产品.

研究的目的:

为了研究两个特定的氨基酸残留在GGPPs的N端螺旋体中的作用.
阐明这些残留物如何通过远程相互作用调节酶二元化和活性.

主要方法:

分析超离心法 (AUC) 用于表征酶二分化.
位点定向的突变发生,以产生特定的氨基酸替代.
分子动力学模拟和自由能量分解分析以了解结构效应.

主要成果:

尽管距离主接口很远,但Saccharomyces cerevisiae GGPPs的N端螺旋体A独特地稳定了二元化.
用Gly取代L8和I9残留物破坏了二元体形成,将其转化为单体.
突变导致GGPPs活动显著减少 (至少是10^3倍).

结论:

由螺旋体A残留L8和I9介导的长距离相互作用对GGPPs二分化和酶功能至关重要.
了解这些相互作用为酶调节和蛋白质结构功能关系提供了洞察力.

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