相关实验视频

Updated: Mar 17, 2026

Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility

Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility

Published on: March 11, 2022

通过反向甘氨基化来表征1家族甘氨基转移酶 (GT1,UGT):快速确定受体特异性,捐赠体特异性,水解和酶稳定性

Mads R Langhorn¹, David Teze^1,2

¹The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark.

|March 16, 2026

概括

开发了一种新的,低成本的高通量选方法,使用逆糖化,用于糖化转移酶 (UGTs). 这种方法有效地描述了UGT及其特异性,使得酶的应用范围更广.

科学领域:

生物化学生物化学
酶学是一种酶学.
化学生物学化学生物学

背景情况:

常规合成的葡萄糖合物是低效的,昂贵的,和不友好的环境.
甘氨基转移酶1家族 (UGTs) 酶为甘氨基化提供了更有效的替代方案,但缺乏有效的选方法.
了解UGT接受器的特异性对于其更广泛的应用至关重要.

研究的目的:

开发一种通用,低成本,高通量选方法,用于UGT的表征.
使用新方法研究UGT动力学,供体特异性和受体特异性.
证明该方法用于测定其他与UGT相关的现象的实用性.

主要方法:

使用2--4-尼托-β-d-糖化物作为通用高通量选工具的反向糖化.
研究了34个UGT的逆糖化动力学和供体特异性 (葡萄糖,银河糖).
对32个UGT进行了44个受体选,确定了11对的K_M参数.

主要成果:

反向糖化法被所有33个测试的UGTs成功催化.
建立了许多UGT的捐赠者特异性概况和受体特异性的特征.
生成了2280个UGT/接受器对的综合数据集,并提供了自动化动力学分析的脚本.

结论:

用2-chloro-4-nitrophenyl-β-d-glycosides进行逆糖化,为高通量UGT测定提供了一个强大的,具有成本效益的和通用的平台.
这种方法显著提高了UGT酶的表征和潜在应用.
开发的选工具和数据集有助于进一步研究UGT的特异性和功能.

更多相关视频

Demonstration of Heterologous Complexes formed by Golgi-Resident Type III Membrane Proteins using Split Luciferase Complementation Assay

Demonstration of Heterologous Complexes formed by Golgi-Resident Type III Membrane Proteins using Split Luciferase Complementation Assay

Published on: September 10, 2020

Identification and Characterization of Protein Glycosylation using Specific Endo- and Exoglycosidases

Identification and Characterization of Protein Glycosylation using Specific Endo- and Exoglycosidases

Published on: December 26, 2011

相关概念视频

Oligosaccharide Assembly

Oligosaccharide Assembly

Protein glycosylation starts in the ER lumen and continues in the Golgi apparatus. Glycosyltransferases catalyze the addition of sugar molecules or glycosylation of proteins. Usually, these enzymes add sugars to the hydroxyl groups of selected serine or threonine residues to form O-linked glycans or the amino groups of asparagine residues to form N-linked glycans. Different positions on the same polypeptide chain can contain differently linked glycans.
Multiple sugar molecules that may or may...

Pharmacogenetics of Phase II Enzymes: N-acetyltransferase, Thiopurine S-methyltransferase, UDP-glucuronosyltransferase

Pharmacogenetics of Phase II Enzymes: N-acetyltransferase, Thiopurine S-methyltransferase, UDP-glucuronosyltransferase

Phase II biotransformation reactions are essential for detoxifying and eliminating xenobiotics, including many pharmaceutical compounds. These reactions typically involve conjugation, the covalent attachment of polar endogenous groups such as glucuronic acid, sulfate, methyl, or acetyl moieties to functional groups introduced during Phase I metabolism. The resulting conjugates are more water-soluble, enabling efficient renal or biliary excretion.The major classes of Phase II enzymes include...

Glucose Transporters

Glucose Transporters

Glucose transporters facilitate the transport of glucose across the cell membrane. In addition to glucose, some glucose transporters can also aid the movement of other hexoses such as fructose, mannose, and galactose.
Facilitated diffusion-glucose transporters (GLUTs) are encoded by the solute-linked carrier (SLC) family 2, subfamily A gene family, or SLC2A. The 14 GLUT protein members are distributed into three classes: