Biochemistry (Moscow)

, Volume 75, Issue 7, pp 938–943 | Cite as

Isoforms of human O-GlcNAcase show distinct catalytic efficiencies

  • Jing LiEmail author
  • Cai-luan Huang
  • Lian-wen Zhang
  • Lin Lin
  • Zhong-hua Li
  • Fu-wu Zhang
  • Peng WangEmail author


O-GlcNAcase (OGA) is a family 84 glycoside hydrolase catalyzing the hydrolytic cleavage of O-linked β-N-acetylglucosamine (O-GlcNAc) from serine and threonine residues of proteins. Thus far, three forms of OGA have been identified in humans. Here we optimized the expression of these isoforms in E. coli and characterized their kinetic properties. Using Geno 3D, we predicted that N-terminal amino acids 63–342 form the catalytic site for O-GlcNAc removal and characterized it. Large differences are observed in the Km value and catalytic efficiency (kcat/Km) for the three OGA variants, though all of them displayed O-GlcNAc hydrolase activity. The full-length OGA had the lowest Km value of 0.26 mM and the highest catalytic efficiency of 3.51·103. These results reveal that the N-terminal region (a.a. 1–350) of OGA contains the catalytic site for glycoside hydrolase and the C-terminal region of the coding sequence has the ability to stabilize the native three-dimensional structure and further affect substrate affinity.

Key words

O-GlcNAcase isoform substrate affinity catalytic efficiency 





full-length O-GlcNAcase




Luria-Bertani (broth)


meningioma expressed antigen 5






programmed cell death


the shortest OGA


variant of OGA


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Copyright information

© Pleiades Publishing, Ltd. 2010

Authors and Affiliations

  1. 1.College of Pharmacy and State Key Laboratory of Element-Organic ChemistryNankai UniversityTianjinChina

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