激素SAM酶酸甲酸酶激活酶的单价离子激活
Krista A Shisler1, Rachel U Hutcheson1, Masaki Horitani2
1Department of Chemistry and Biochemistry, Montana State University , Bozeman, Montana 59717, United States.
Journal of the American Chemical Society
|August 4, 2017
概括
酸甲酸酶激活酶 (PFL-AE) 需要单价离子来产生活性. 这种称为K+的阳离子通过与S-adenosyl-l-methionine (SAM) 和铁-硫团相互作用来优化酶功能.
科学领域:
- 生物化学
- 酵素学
- 结构生物学
背景情况:
- 酸甲酸酶激活酶 (PFL-AE) 对于生成酸甲酸甲酸酶中的糖基至关重要.
- 激进的S-adenosyl-l-methionine (SAM) 酶与维生素B12依赖的酶具有机理上的相似性.
研究的目的:
- 描述PFL-AE中的单价离子结合点.
- 阐明这种离子在酶活性和机制中的作用.
- 为了研究阴离子与SAM辅基质和铁硫集群的相互作用.
主要方法:
- 用X射线结晶学进行结构测定.
- 脉冲电子核双共振 (ENDOR) 光谱用于溶液状态分析.
- 用于电子性能评估的电子磁共振 (EPR) 和循环二极化 (CD) 光谱仪.
- 生物化学测定以评估各种的酶活性.
主要成果:
- 在其活性位点上,PFL-AE结合了一种催化必需的单价离子 (M+),在晶体结构中被确定为Na+和溶液中为23Na.
- 阴离子与SAM碳酸盐氧相互作用,并影响SAM化铁硫团的电子性质.
- 酶活性高度依赖于阳离子类型,观察到K+和NH4+的最佳活性.
结论:
- PFL-AE是一种I型M+激活酶,其中通过与SAM的相互作用来调节反应性.
- 的存在和身份对于PFL-AE的催化功能至关重要.
- 这种依赖阴离子的机制为SAM激素酶调节提供了新的见解.
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