caa a22 4500 445861517 CHVBK 20180317145444.0 cr unu---uuuuu 170323e20110301xx s 000 0 eng 10.1007/s00439-010-0914-4 doi (NATIONALLICENCE)springer-10.1007/s00439-010-0914-4 Insight into the biochemical characteristics of a novel glucokinase gene mutation [Elektronische Daten] [Yunfeng Shen, Mengyin Cai, Hua Liang, Hongwei Wang, Jianping Weng] 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. Springer-Verlag, 2010 Shen Yunfeng Department of Endocrinology, The Third Affiliated Hospital of Sun Yat-sen University, Research Center for Diabetes Care of Guangdong Province, 600 Tianhe Road, 510630, Guangzhou, Guangdong, People's Republic of China aut Cai Mengyin Department of Endocrinology, The Third Affiliated Hospital of Sun Yat-sen University, Research Center for Diabetes Care of Guangdong Province, 600 Tianhe Road, 510630, Guangzhou, Guangdong, People's Republic of China aut Liang Hua Department of Endocrinology, The Third Affiliated Hospital of Sun Yat-sen University, Research Center for Diabetes Care of Guangdong Province, 600 Tianhe Road, 510630, Guangzhou, Guangdong, People's Republic of China aut Wang Hongwei Section of Endocrinology, Department of Medicine, University of Chicago, 5841 S. Maryland Ave, 60637, Chicago, IL, USA aut Weng Jianping Department of Endocrinology, The Third Affiliated Hospital of Sun Yat-sen University, Research Center for Diabetes Care of Guangdong Province, 600 Tianhe Road, 510630, Guangzhou, Guangdong, People's Republic of China aut Human Genetics Springer-Verlag 129/3(2011-03-01), 231-238 0340-6717 129:3<231 2011 129 439 https://doi.org/10.1007/s00439-010-0914-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00439-010-0914-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Shen Yunfeng Department of Endocrinology, The Third Affiliated Hospital of Sun Yat-sen University, Research Center for Diabetes Care of Guangdong Province, 600 Tianhe Road, 510630, Guangzhou, Guangdong, People's Republic of China aut NATIONALLICENCE 700 1- Cai Mengyin Department of Endocrinology, The Third Affiliated Hospital of Sun Yat-sen University, Research Center for Diabetes Care of Guangdong Province, 600 Tianhe Road, 510630, Guangzhou, Guangdong, People's Republic of China aut NATIONALLICENCE 700 1- Liang Hua Department of Endocrinology, The Third Affiliated Hospital of Sun Yat-sen University, Research Center for Diabetes Care of Guangdong Province, 600 Tianhe Road, 510630, Guangzhou, Guangdong, People's Republic of China aut NATIONALLICENCE 700 1- Wang Hongwei Section of Endocrinology, Department of Medicine, University of Chicago, 5841 S. Maryland Ave, 60637, Chicago, IL, USA aut NATIONALLICENCE 700 1- Weng Jianping Department of Endocrinology, The Third Affiliated Hospital of Sun Yat-sen University, Research Center for Diabetes Care of Guangdong Province, 600 Tianhe Road, 510630, Guangzhou, Guangdong, People's Republic of China aut NATIONALLICENCE 773 0- Human Genetics Springer-Verlag 129/3(2011-03-01), 231-238 0340-6717 129:3<231 2011 129 439 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer