相关实验视频

Updated: Jul 27, 2025

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

Published on: December 12, 2017

对阿尔多斯降解酶的突变研究,以追踪一个短暂的口袋开口,并解释干亲属性悬崖

Lea-Sophie Klee¹, Marina Gárdonyi¹, Tobias Hüfner¹

¹Institute of Pharmaceutical Chemistry, Philipps University Marburg, Marbacher Weg 6, 35032, Marburg, Germany.

|June 6, 2023

概括

针对糖尿病并发症的人类阿尔多缩酶抑制剂显示结合的改变. 突变的守门者残留物影响抑制剂亲和力和口袋的开放,揭示了对阿尔多减少酶药物设计的见解.

科学领域:

生物化学生物化学
药理学药理学是指药理学的学科.
分子生物学分子生物学

背景情况:

人类阿尔多缩酶是预防糖尿病并发症的关键目标.
该酶拥有一个短暂的特异性口袋,在结合强效抑制剂时会打开.
守门的残留物控制着这个口袋的打开.

研究的目的:

为了研究人类阿尔多斯减少酶中暂时口袋开放的机制.
了解守门残留物中的突变如何影响抑制剂结合.
阐明在酶-抑制剂相互作用中的连接物溶解和 conformational 选择的作用.

主要方法:

在守门机制中的氨酸残留物对氨酸的局部定向突变发生.
使用两种具有和碳氧基团的同结构抑制剂的结合亲和性研究.
对连接体和口袋溶解性质的分析.
诱导适应与构造性选择模型的评估.

主要成果:

突变的守门者残留物减少了两种抑制剂之间的结合亲和力差异,从1000倍到10倍.
基衍生抑制剂失去了亲和力,但在突变变体中保留了与开口口袋的结合.
碳酸盐模拟物显示出最小的亲和力变化,但结合偏好从封闭到开放的口袋状态的转变.
改变的连接体和口袋溶解,以及从诱导适合转向形状选择的转变解释了观察到的结合变化.

结论:

守门残留物在调节阿尔多减少酶暂时口袋的开放方面发挥着至关重要的作用.
连接物溶解和 conformational 选择是影响抑制剂结合动态的关键因素.
了解这些机制可以指导对阿尔多减少酶抑制剂用于糖尿病并发症的合理设计.

关键词:

其他ALR2变体亲密关系悬崖的悬崖.开放机制的开放机制.一个短暂的口袋.供水网络的水道网络.

更多相关视频

Deciphering the Structural Effects of Activating EGFR Somatic Mutations with Molecular Dynamics Simulation

Deciphering the Structural Effects of Activating EGFR Somatic Mutations with Molecular Dynamics Simulation

Published on: May 20, 2020

Achieving Efficient Fragment Screening at XChem Facility at Diamond Light Source

Achieving Efficient Fragment Screening at XChem Facility at Diamond Light Source

Published on: May 29, 2021

相关概念视频

Ligand Binding and Linkage

Ligand Binding and Linkage

Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked. In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...

Allosteric Proteins-ATCase

Allosteric Proteins-ATCase

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...

Cooperative Allosteric Transitions

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...

Ligand Binding Sites

Ligand Binding Sites

Proteins are dynamic macromolecules that carry out a wide variety of essential processes; however, the activities of most proteins depend on their interactions with other molecules or ions, known as ligands.
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...

The Equilibrium Binding Constant and Binding Strength

The Equilibrium Binding Constant and Binding Strength

The equilibrium binding constant (Kb) quantifies the strength of a protein-ligand interaction. Kb can be calculated as follows when the reaction is at equilibrium:

Conserved Binding Sites

Conserved Binding Sites

Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...