6TKA

Crystal structure of human O-GlcNAc transferase bound to substrate 7 and a peptide from HCF-1 pro-repeat 2 (11-26)

これはPDB形式変換不可エントリーです。

6TKA の概要

• エントリーDOI: 10.2210/pdb6tka/pdb
• 分子名称: UDP-N-acetylglucosamine--peptide N-acetylglucosaminyltransferase 110 kDa subunit, HCF-1 pro-repeat 2 (11-26), [[(2~{R},3~{S},4~{R},5~{R})-5-[2,4-bis(oxidanylidene)pyrimidin-1-yl]-3,4-bis(oxidanyl)oxolan-2-yl]methoxy-oxidanyl-phosphoryl] [(2~{R},3~{R},4~{R},5~{S},6~{R})-3-[6-(butanoylamino)hexanoylamino]-6-(hydroxymethyl)-4,5-bis(oxidanyl)oxan-2-yl] hydrogen phosphate, ... (5 entities in total)
• 機能のキーワード: transferase complex o-glcnac ogt, transferase
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 84066.30

構造登録者

Meek, R.W.,Davies, G.J. (登録日: 2019-11-28, 公開日: 2020-03-04, 最終更新日: 2024-01-24)

主引用文献

Alteen, M.G.,Gros, C.,Meek, R.W.,Cardoso, D.A.,Busmann, J.A.,Sangouard, G.,Deen, M.C.,Tan, H.Y.,Shen, D.L.,Russell, C.C.,Davies, G.J.,Robinson, P.J.,McCluskey, A.,Vocadlo, D.J.
A Direct Fluorescent Activity Assay for Glycosyltransferases Enables Convenient High-Throughput Screening: Application to O-GlcNAc Transferase.
Angew.Chem.Int.Ed.Engl., 59:9601-9609, 2020

PubMed Abstract: Glycosyltransferases carry out important cellular functions in species ranging from bacteria to humans. Despite their essential roles in biology, simple and robust activity assays that can be easily applied to high-throughput screening for inhibitors of these enzymes have been challenging to develop. Herein, we report a bead-based strategy to measure the group-transfer activity of glycosyltransferases sensitively using simple fluorescence measurements, without the need for coupled enzymes or secondary reactions. We validate the performance and accuracy of the assay using O-GlcNAc transferase (OGT) as a model system through detailed Michaelis-Menten kinetic analysis of various substrates and inhibitors. Optimization of this assay and application to high-throughput screening enabled screening for inhibitors of OGT, leading to a novel inhibitory scaffold. We believe this assay will prove valuable not only for the study of OGT, but also more widely as a general approach for the screening of glycosyltransferases and other group-transfer enzymes.

PubMed: 32092778
DOI: 10.1002/anie.202000621

実験手法

X-RAY DIFFRACTION (1.91 Å)