相关实验视频

Updated: Aug 8, 2026

Production, Crystallization and Structure Determination of C. difficile PPEP-1 via Microseeding and Zinc-SAD

Production, Crystallization and Structure Determination of C. difficile PPEP-1 via Microseeding and Zinc-SAD

Published on: December 30, 2016

对Clostridioides difficile endolysin Ecd09610葡萄糖氨基酶域的X射线结构和突变发生分析

Hiroshi Sekiya¹, Yasuhiro Nonaka², Shigehiro Kamitori³

¹Department of Infectious Disease, College of Pharmaceutical Science, Matsuyama University, 4-2 Bunkyo-cho, Matsuyama, Ehime, 790-8578, Japan.

Biochemical and biophysical research communications

|April 30, 2024

概括

克洛斯特里迪奥伊德难度内素 (Ecd09610) 有一个葡萄糖胺酶域 (Ecd09610CD53),其结构保留在GH73酶中. 突变酶确定了关键活性残留物,并进行比较,发现其活性低于Clostridium perfringens autolysin (AcpCD).

科学领域:

酶学是一种酶学.
结构生物学结构生物学
微生物学微生物学

背景情况:

克洛斯特里迪奥伊德 (Clostridioides difficile endolysin) (Ecd09610) 具有N端未知和C端催化域 (葡萄糖胺酶和内酶).
葡萄糖氨基酶域 (Ecd09610CD53) 对酶活性至关重要.

研究的目的:

为了阐明Clostridioides difficile内素葡萄糖胺酶域的结构和功能 (Ecd09610CD53).
为了确定Ecd09610CD53的催化活性所必需的关键残留物.
为了比较Ecd09610CD53与其他GH73家族酶的活性.

主要方法:

使用X射线晶体学来确定Ecd09610CD53.5的结构.
用定位突变发生法来研究特定残留物的作用.
进行了细胞图分析以进行活动比较.

主要成果:

Ecd09610CD53表现出具有9个螺旋体的α-捆状结构,这是GH73家族酶的特征.
谷氨酸405 (Glu405) 和氨酸470 (Asn470) 被确定为对葡萄糖氨基酶活性必不可少的.
一个涉及Glu405和Asn470的邻群机制被提出用于催化.
克洛斯特里透的自解酶催化域 (AcpCD) 显示出比Ecd09610CD53.5高15倍的活性.
基质结合槽深度与GH73家族的葡萄糖氨基酶活性强度相关.

结论:

Ecd09610CD53的结构和功能特征为GH73家族酶机制提供了洞察力.
Glu405和Asn470对于Ecd09610CD53.3的催化功能至关重要.
酶结构的变化,特别是基质结合槽深度,影响葡萄糖胺酶活性.

关键词:

克洛斯特里迪奥伊德斯困难菌恩多利辛 (Endolysin) 是一种内分泌物.葡萄糖氨基酸酶的使用突变分析突变分析在X射线中,X射线结构结构是什么?

更多相关视频

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

Published on: February 5, 2019

Structural Biology and Analytical Chemistry Approaches for Characterizing C-Glycoside Metabolic Enzymes in Human Gut Microbiota

Structural Biology and Analytical Chemistry Approaches for Characterizing C-Glycoside Metabolic Enzymes in Human Gut Microbiota

Published on: May 23, 2025

相关概念视频

$X-ray Diffraction of Biological Samples$ 01:10

X-ray Diffraction of Biological Samples

X-ray diffraction or XRD is an analytical tool that utilizes X-rays to study ordered structures such as crystalline organic and inorganic samples, polycrystalline materials, proteins, carbohydrates, and drugs.
According to Bragg's law, when X-rays strike the sample positioned on a stage, the rays are scattered by the electron clouds around the sample atoms. The X-ray diffraction or scattering is caused by constructive interference of the X-ray waves that reflect off the internal crystal...