Microscale thermophoresis as a powerful tool for screening glycosyltransferases involved in cell wall biosynthesis.

ABSTRACT:

Background

Identification and characterization of key enzymes associated with cell wall biosynthesis and modification is fundamental to gain insights into cell wall dynamics. However, it is a challenge that activity assays of glycosyltransferases are very low throughput and acceptor substrates are generally not available.

Results

We optimized and validated microscale thermophoresis (MST) to achieve high throughput screening for glycosyltransferase substrates. MST is a powerful method for the quantitative analysis of protein-ligand interactions with low sample consumption. The technique is based on the motion of molecules along local temperature gradients, measured by fluorescence changes. We expressed glycosyltransferases as YFP-fusion proteins in tobacco and optimized the M

SUBMITTER: Shao W 

PROVIDER: S-EPMC7389378 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature