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Insight into the biochemical characteristics of a novel glucokinase gene mutation

Abstract

Glucokinase (GCK) acts as a glucose sensor and regulates β-cell insulin secretion. The heterozygous mutations in the gene encoding GCK cause a reduction of the enzyme activity, which results in a monogenic form of diabetes, maturity-onset diabetes of the young. In the present study, we identified and functionally characterized a novel missense mutation in the GCK gene, which results in a protein mutation Glu339 → Lys (E339K), from a Chinese family with hyperglycemia. The same GCK mutation that co-segregated with diabetes phenotype was identified in five members of this family but was not found in 200 healthy control individuals. We expressed and affinity-purified the GCK proteins from bacterial expression system that carries mutation (E339K) and fused to glutathione S-transferase. The expressed GCK protein was subjected to the measurement of its biochemical effects of the missense mutation on GCK activity. Our results showed that the mutation reduced the GCK protein yield. The enzymatic kinetics and the thermal stability analysis on the recombinant GCK proteins revealed that the mutation inactivates enzyme kinetics and severely impaired the GCK protein stability.

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Acknowledgments

We thank the family in this study. Professors Franz M. Matschinsky, Pan Chen, and Charles A. Stanley are thanked for generously providing the wild-type GST-GCK-cDNA. This work was supported by Grants from the National Basic Research Program of China (2006CB503902), the Ministry of Health of the People’s Republic of China for Key Projects of Clinical Disciplines of Hospitals Affiliated to Ministry of Health (2007–2009), the Specialized Research Fund for the Doctoral Program of Higher Education of China (200805580070) to WJ, and Post-doctor Foundation of Ministry of Education (20060390754), Natural Science Foundation of Guangdong Province, China (8151008901000196), Medical Science Foundation of Guangdong Province, China (B2008045) to LH.

Conflict of interest

None.

Author information

Correspondence to Jianping Weng.

Additional information

Y. Shen and M. Cai contributed equally to this work.

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Shen, Y., Cai, M., Liang, H. et al. Insight into the biochemical characteristics of a novel glucokinase gene mutation. Hum Genet 129, 231–238 (2011). https://doi.org/10.1007/s00439-010-0914-4

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Keywords

  • Gestational Diabetes Mellitus
  • Oral Glucose Tolerance Test
  • Thermal Instability
  • E339K Mutation
  • Glutamic Acid Decarboxylase Antibody