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一个乙转移酶 ribozyme 的结构和运动特征
H Suga1, P A Lohse, J W Szostak
1Department of Molecular Biology, Massachusetts General Hospital, Boston 02114, USA.
Journal of the American Chemical Society
|September 7, 2001
概括
研究人员设计了一种用于乙转移的 ribozyme,通过突变证实了它的结构和功能. 关键G:U波动对通过稳定整个 ribozyme 结构内的基质结合来增强催化作用.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 化学生物学 化学生物学
背景情况:
- 乙转移酶 рибо酶对于催化乙基转移至关重要.
- 实验室内选择已被用来进化具有特定催化功能的新型 ribozymes.
- 了解 ribozyme 结构-功能关系是它们在生物技术中的应用的关键.
研究的目的:
- 为了阐明在体外选择的乙转移酶 ribozyme 的二次结构.
- 调查特定结构元素的作用,包括G:U摇摆对,在 ribozyme 催化中.
- 描述工程化 рибо酶的基质结合和催化机制.
主要方法:
- 在体外选择以分离乙转移酶 ribozyme 变体.
- 进化的核糖酶的序列分析,以预测二级结构.
- 定位基因突变 (单一和补偿双重突变) 用于测试结构模型.
- 野生型和突变型 ribozymes 的动态特征.
- 竞争性抑制试验用于探测基质结合相互作用.
主要成果:
- 为乙转移酶 рибо酶生成了一种二级结构模型,并经过实验验证.
- 模板域被证实对于调整基质和催化区域至关重要.
- 在模板域中发现两个并列的G:U波动基对可提高催化速率.
- 这些摇摆对的催化贡献取决于上下文,需要完整的 ribozyme 结构.
- 核酶调节金属结合特性在摇摆对位点.
- 基质结合被 ribozyme 模板稳定,而氨基酸相互作用会破坏结合的稳定.
结论:
- 该研究成功确定并验证了工程化乙转移酶 ribozyme 的二次结构.
- 特定的结构特征,特别是G:U摇摆对,对于高效的催化和基质结合至关重要.
- 这些发现提供了对 ribozyme 中介的转移和基质识别机制的见解.
相关概念视频
Enzyme Kinetics
Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
Riboswitches
Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
Ribozymes
The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
Ribozymes can be...
Introduction to Mechanisms of Enzyme Catalysis
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 a mild...
Ribozymes
The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
Ribozymes can be...
Introduction to Mechanisms of Enzyme Catalysis
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 a mild...

