Expression of glucokinase in glucose-unresponsive human fetal pancreatic islet-like cell clusters

Jian Tu, Bernard E. Tuch*

*Corresponding author for this work

    Research output: Contribution to journalArticle

    11 Citations (Scopus)

    Abstract

    Glucokinase (GK) is the glucose sensor in the adult β-cell, resulting in fuel for insulin synthesis and secretion. Defects in this enzyme in the β-cell are responsible for the genetic disorder maturity-onset diabetes of the young, with the β-cell being unable to secrete insulin appropriately when challenged with glucose. The human fetal β-cell is also unable to secrete insulin when exposed to glucose, but whether GK is present and functional in this developing cell is unknown. To determine the expression of GK in human fetal pancreatic tissue, cytosolic protein was extracted from human fetal islet-like cell clusters (ICCs) at 17-19 weeks gestation and examined for protein content and enzyme activity. On Western blots, a single band corresponding to GK was seen at 52 kDa, and this was similar to that obtained from human adult islets. The maximal velocity (V(max)) of GK was less in fetal ICCs than that in adult islets (8.7 vs. 20.7 nmol/mg protein · h); similar K(m) values were found in both ICCs and islets. No attempt was made to determine which cells in an ICC contained GK. Glucose utilization was determined radiometrically; the V(max) of the high K(m) component was less in ICCs than in islets (31.3 pmol/ICC · h vs. 101.4 pmol/islet · h). Culture of ICCs for 3-7 days in medium containing 11.2 mmol/L glucose resulted in a 3.7-fold increase in the V(max) of GK and a 1.8-fold increase in glucose utilization. These enhanced activities of glucose phosphorylation and glycolysis, however, did not lead to the β-cell being able to secrete insulin when exposed to glucose. In conclusion, glucokinase is present and functional in human fetal ICCs, but the inability of the human fetal/β-cell to secrete insulin in response to an acute glucose challenge is not due to immaturity of this enzyme.

    Original languageEnglish
    Pages (from-to)943-948
    Number of pages6
    JournalJournal of Clinical Endocrinology and Metabolism
    Volume82
    Issue number3
    DOIs
    Publication statusPublished - 1997

    Fingerprint Dive into the research topics of 'Expression of glucokinase in glucose-unresponsive human fetal pancreatic islet-like cell clusters'. Together they form a unique fingerprint.

  • Cite this