2J62

Structure of a bacterial O-glcnacase in complex with glcnacstatin

Summary for 2J62

• Entry DOI: 10.2210/pdb2j62/pdb
• Descriptor: O-GlcNAcase NagJ, CHLORIDE ION, N-[(5R,6R,7R,8S)-6,7-DIHYDROXY-5-(HYDROXYMETHYL)-2-(2-PHENYLETHYL)-1,5,6,7,8,8A-HEXAHYDROIMIDAZO[1,2-A]PYRIDIN-8-YL]-2-METHYLPROPANAMIDE, ... (4 entities in total)
• Functional Keywords: hydrolase
• Biological source: Clostridium perfringens
• Total number of polymer chains: 2
• Total formula weight: 134270.11

Authors

Dorfmueller, H.C.,Borodkin, V.S.,Schimpl, M.,Shepherd, S.M.,Shpiro, N.A.,van Aalten, D.M.F. (deposition date: 2006-09-22, release date: 2007-02-13, Last modification date: 2024-05-08)

Primary citation

Dorfmueller, H.C.,Borodkin, V.S.,Schimpl, M.,Shepherd, S.M.,Shpiro, N.A.,van Aalten, D.M.
GlcNAcstatin: a picomolar, selective O-GlcNAcase inhibitor that modulates intracellular O-glcNAcylation levels.
J. Am. Chem. Soc., 128:16484-16485, 2006

PubMed Abstract: Many phosphorylation signal transduction pathways in the eukaryotic cell are modulated by posttranslational modification of specific serines/threonines with N-acetylglucosamine (O-GlcNAc). Levels of O-GlcNAc on key proteins regulate biological processes as diverse as the cell cycle, insulin signaling, and protein degradation. The two enzymes involved in this dynamic and abundant modification are the O-GlcNAc transferase and O-GlcNAcase. Structural data have recently revealed that the O-GlcNAcase possesses an active site with significant structural similarity to that of the human lysosomal hexosaminidases HexA/HexB. PUGNAc, an O-GlcNAcase inhibitor widely used to raise levels of O-GlcNAc in human cell lines, also inhibits these hexosaminidases. Here, we have exploited recent structural information of an O-GlcNAcase-PUGNAc complex to design and synthesize a glucoimidazole-based inhibitor, GlcNAcstatin, which is a 5 pM competitive inhibitor of enzymes of the O-GlcNAcase family, shows 100000-fold selectivity over HexA/B, and binds to the O-GlcNAcase active site by mimicking the transition state as revealed by X-ray crystallography. This compound is able to raise O-GlcNAc levels in human HEK 293 and SH-SY5Y neuroblastoma cell lines and thus provides a novel, potent tool for the study of the role of O-GlcNAc in intracellular signal transduction pathways.

PubMed: 17177381
DOI: 10.1021/ja066743n

Experimental method

X-RAY DIFFRACTION (2.26 Å)