Human Genetics

, Volume 129, Issue 3, pp 231–238

Insight into the biochemical characteristics of a novel glucokinase gene mutation

  • Yunfeng Shen
  • Mengyin Cai
  • Hua Liang
  • Hongwei Wang
  • Jianping Weng
Original Investigation

DOI: 10.1007/s00439-010-0914-4

Cite this article as:
Shen, Y., Cai, M., Liang, H. et al. Hum Genet (2011) 129: 231. doi:10.1007/s00439-010-0914-4

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.

Supplementary material

439_2010_914_MOESM1_ESM.doc (68 kb)
Supplementary material 1 (DOC 67.5 kb)

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Yunfeng Shen
    • 1
    • 3
  • Mengyin Cai
    • 1
  • Hua Liang
    • 1
  • Hongwei Wang
    • 2
  • Jianping Weng
    • 1
  1. 1.Department of Endocrinology, The Third Affiliated Hospital of Sun Yat-sen UniversityResearch Center for Diabetes Care of Guangdong ProvinceGuangzhouPeople’s Republic of China
  2. 2.Section of Endocrinology, Department of MedicineUniversity of ChicagoChicagoUSA
  3. 3.Department of Endocrinology and MetabolismThe Second Affiliated Hospital of Nanchang UniversityNanchangPeople’s Republic of China

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