5VIF

Electrophilic probes for deciphering substrate recognition by O-GlcNAc transferase

5VIF の概要

• エントリーDOI: 10.2210/pdb5vif/pdb
• 関連するPDBエントリー: 5VIE
• 分子名称: UDP-N-acetylglucosamine--peptide N-acetylglucosaminyltransferase 110 kDa subunit, CKII, URIDINE-5'-DIPHOSPHATE, ... (5 entities in total)
• 機能のキーワード: o-glcnac transferase, glycosylation electrophilic probes, ckii, 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: 83058.92

構造登録者

Jiang, J.,Li, B.,Hu, C.-W.,Worth, M.,Fan, D.,Li, H. (登録日: 2017-04-15, 公開日: 2017-10-18, 最終更新日: 2024-10-23)

主引用文献

Hu, C.W.,Worth, M.,Fan, D.,Li, B.,Li, H.,Lu, L.,Zhong, X.,Lin, Z.,Wei, L.,Ge, Y.,Li, L.,Jiang, J.
Electrophilic probes for deciphering substrate recognition by O-GlcNAc transferase.
Nat. Chem. Biol., 13:1267-1273, 2017

PubMed Abstract: O-linked β-N-acetylglucosamine (O-GlcNAc) transferase (OGT) is an essential human glycosyltransferase that adds O-GlcNAc modifications to numerous proteins. However, little is known about the mechanism with which OGT recognizes various protein substrates. Here we report on GlcNAc electrophilic probes (GEPs) to expedite the characterization of OGT-substrate recognition. Data from mass spectrometry, X-ray crystallization, and biochemical and radiolabeled kinetic assays support the application of GEPs to rapidly report the impacts of OGT mutations on protein substrate or sugar binding and to discover OGT residues crucial for protein recognition. Interestingly, we found that the same residues on the inner surface of the N-terminal domain contribute to OGT interactions with different protein substrates. By tuning reaction conditions, a GEP enables crosslinking of OGT with acceptor substrates in situ, affording a unique method to discover genuine substrates that weakly or transiently interact with OGT. Hence, GEPs provide new strategies to dissect OGT-substrate binding and recognition.

PubMed: 29058723
DOI: 10.1038/nchembio.2494

実験手法

X-RAY DIFFRACTION (2.25 Å)