环侧链在三酸盐异构酶催化中的作用
Xiang Zhai1, Tina L Amyes1, John P Richard1
1Department of Chemistry, University at Buffalo, SUNY , Buffalo, New York 14260-3000, United States.
Journal of the American Chemical Society
|November 17, 2015
概括
三酸盐异构酶 (TIM) 的突变会影响酶催化. 大多数突变会类似地改变基质结合和过渡状态,而一些突变会影响催化位的反应性.
科学领域:
- 生物化学
- 酶动力学
- 蛋白质工程
背景情况:
- 三酸盐异构酶 (TIM) 通过键稳定酶基质复合物.
- 在TIM的循环7中的特定残留物 (Y208,S211) 与循环6相互作用.
- 了解这些相互作用是酶功能和工程的关键.
研究的目的:
- 研究Y208和S211突变对TIM运动参数的影响.
- 分析这些突变如何影响过渡状态的稳定.
- 阐明基质结合能和催化效率之间的关系.
主要方法:
- 在Y208和S211的TIM定位突变.
- 测量整个基板 (DHAP,GAP) 和基板片段 (糖甲,酸) 的动态参数.
- 动力数据的线性对比分析.
主要成果:
- 大多数TIM突变对整个基板和基板片段的激活障碍产生了类似的变化.
- 在催化速率 (kcat) 和过渡状态结合能量 (Kd) 之间观察到相关性.
- Y208突变主要降低了离子结合能量,而Y208F影响了催化部位.
结论:
- TIM突变通常会对过渡状态稳定产生均影响.
- 酶基质结合能量在迈凯利斯复合体和过渡状态之间进行分离.
- 特定的突变可以改变除了简单的结合能效之外的催化位点反应性.
相关概念视频
Catalytically Perfect Enzymes
5.4K
The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at high-speed. Their catalytic efficiency values range from 108-109 M-1s-1. These enzymes are also called 'diffusion-controlled' as the only rate-limiting step in the catalysis is that of the substrate diffusion into the active site. Examples include triose phosphate isomerase, fumarase, and superoxide dismutase.
Most enzymes...
Most enzymes...
5.4K
ATP Synthase: Mechanism
18.8K
In animals, the mitochondrial F1F0 ATP synthase is the key protein that synthesizes ATP molecules through a complex catalytic mechanism. While the nuclear genome encodes the majority of ATP synthase subunits, the mitochondrial genome encodes some of the enzyme's most critical components. The formation of this multi-subunit enzyme is a complex multi-step process regulated at the level of transcription, translation, and assembly. Defects in one or more of these steps can result in decreased...
18.8K
Introduction to Mechanisms of Enzyme Catalysis
10.2K
10.2K
Introduction to Mechanisms of Enzyme Catalysis
11.4K
For many years, scientists thought that enzyme-substrate binding took place in a simple "lock-and-key" fashion. This model stated that the enzyme and substrate fit together perfectly in one instantaneous step. However, current research supports a more refined view scientists call induced fit. The induced-fit model expands upon the lock-and-key model by describing a more dynamic interaction between enzyme and substrate. As the enzyme and substrate come together, their interaction causes...
11.4K
Allosteric Proteins-ATCase
6.9K
Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...
6.9K
ATP Synthase: Structure
17.5K
ATP synthase or ATPase is among the most conserved proteins found in bacteria, mammals, and plants. This enzyme can catalyze a forward reaction in response to the electrochemical gradient, producing ATP from ADP and inorganic phosphate. ATP synthase can also work in a reverse direction by hydrolyzing ATP and generating an electrochemical gradient. Different forms of ATP synthases have evolved special features to meet the specific demands of the cell. Based on their specific feature, ATP...
17.5K


