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Isoforms of human O-GlcNAcase show distinct catalytic efficiencies

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An Errata to this article was published on 27 October 2010

Abstract

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.

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Abbreviations

DTT:

dithiothreitol

fOGA:

full-length O-GlcNAcase

IPTG:

isopropyl-L-thio-β-D-galactopyranoside

LB:

Luria-Bertani (broth)

MGEA5:

meningioma expressed antigen 5

4-MU-GlcNAc:

4-methylumbelliferyl-2-acetamido-2-deoxy-β-D-glucopyranoside

OGA:

O-GlcNAcase

PCD:

programmed cell death

sOGA:

the shortest OGA

vOGA:

variant of OGA

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Authors and Affiliations

  1. College of Pharmacy and State Key Laboratory of Element-Organic Chemistry, Nankai University, Tianjin, 300071, China

    Jing Li, Cai-luan Huang, Lian-wen Zhang, Lin Lin, Zhong-hua Li, Fu-wu Zhang & Peng Wang

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  1. Jing Li
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  2. Cai-luan Huang
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  3. Lian-wen Zhang
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  4. Lin Lin
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  5. Zhong-hua Li
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  6. Fu-wu Zhang
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  7. Peng Wang
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Correspondence to Jing Li or Peng Wang.

Additional information

An erratum to this article can be found at http://dx.doi.org/10.1134/S0006297910100160

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Li, J., Huang, Cl., Zhang, Lw. et al. Isoforms of human O-GlcNAcase show distinct catalytic efficiencies. Biochemistry Moscow 75, 938–943 (2010). https://doi.org/10.1134/S0006297910070175

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  • DOI: https://doi.org/10.1134/S0006297910070175

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