相关实验视频

Updated: Sep 14, 2025

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes

Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes

Published on: April 4, 2025

对特定突变影响Taq聚合酶动力学和结构的分子洞察力:一个分子动力学模拟研究

Seddigheh Borhani¹, Seyed Shahriar Arab²

¹Biophysics Department, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.

Journal of biomolecular structure & dynamics

|July 25, 2025

概括

研究人员设计了Taq DNA聚合酶 (Taq pol) 突变,以提高酶的稳定性. 与野生类型酶相比,特定的突变TaqG389E和一个五个突变的变体显示出减少的灵活性和改善的结构完整性.

科学领域:

生物技术是生物技术.
酵素工程是什么? 酶工程是什么
结构生物学结构生物学

背景情况:

Taq DNA聚合酶 (Taq pol) 对生物技术至关重要,但需要增强的热稳定性.
目前的方法还没有完全满足对更稳定的Taq pol变体的需求.

研究的目的:

在Taq pol中进行工程突变,以提高其结构稳定性.
研究这些突变对酶动态和相互作用的影响.

主要方法:

采用基于序列和基于结构的合理设计来进行突变发生.
在Taq pol结构中引入了五种特定突变.
利用分子动力学模拟来分析突变酶结构和动力学.

主要成果:

所有突变酶都显示出更紧的结构,内部键和二次结构含量增加.
突变TaqG389E和TaqS290K arrayA353K arrayL365E arrayN384E arrayG389E表现出最少的结构动态. 突变TaqG389E和TaqS290K arrayA353K arrayL365E arrayN384E表现出最少的结构动态.
突变通常增强非结合性相互作用,并对整体结构稳定性产生积极影响.

结论:

设计的突变成功地提高了Taq的结构稳定性.
突变TaqG389E和TaqS290K robiA353K robiL365E robiN384E的TaqG389E比野生类型的Taq pol更稳定,因为灵活性降低和相互作用增强.

关键词:

在Taq聚合酶.在形设计中.分子动力学模拟模拟蛋白质工程工程蛋白质工程结构稳定性结构稳定性

更多相关视频

Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches

Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches

Published on: October 13, 2022

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

Published on: October 6, 2017

相关概念视频

Translesion DNA Polymerases

Translesion DNA Polymerases

Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...

Proofreading

Proofreading

Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore, it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase...

Spontaneous and Induced Mutations

Spontaneous and Induced Mutations

Spontaneous mutations arise infrequently during DNA replication due to errors in the process. A key factor behind these errors is tautomeric shifts in nitrogenous bases, where bases transition from keto to enol forms or amino to imino forms. This shift can alter base-pairing rules, leading to mutations. Additionally, reactive oxygen species (ROS) arising from aerobic metabolism can damage DNA, resulting in depurination (loss of a purine base) or depyrimidination (loss of a pyrimidine base).