A novel glucokinase regulator in pancreatic beta cells: precursor of propionyl-CoA carboxylase beta subunit interacts with glucokinase and augments its activity.
A glucokinase regulatory protein has been reported to exist in the liver, which suppresses enzyme activity in a complex with fructose 6-phosphate, whereas no corresponding protein has been found in pancreatic beta cells. To search for such a protein in pancreatic beta cells, we screened for a cDNA library of the HIT-T15 cell line with the cDNA of glucokinase from rat islet by the yeast two hybrid system. We detected a cDNA encoding the precursor of propionyl-CoA carboxylase beta subunit (pbetaPCCase), and glutathione S-transferase pull-down assay illustrated that pbetaPCCase interacted with recombinant rat islet glucokinase and with glucokinase in rat liver and islet extracts. Functional analysis indicated that pbetaPCCase decreased the K(m) value of recombinant islet glucokinase for glucose by 18% and increased V(max) value by 23%. We concluded that pbetaPCCase might be a novel activator of glucokinase in pancreatic beta cells.
Shiraishi
A
A
Department of Metabolism and Clinical Nutrition, Graduate School of Medicine, Kyoto University, Sakyoku, Japan. siraisi@metab.kuhp.kyoto-u.ac.jp
Yamada
Y
Y
Tsuura
Y
Y
Fijimoto
S
S
Tsukiyama
K
K
Mukai
E
E
Toyoda
Y
Y
Miwa
I
I
Seino
Y
Y
D000818:Animals; D019735:Carbon-Carbon Ligases / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D015723:Gene Library; D005941:Glucokinase / Q000235:genetics / Q000378:metabolism*; D007515:Islets of Langerhans / Q000378:metabolism*; D008297:Male; D011485:Protein Binding; D011498:Protein Precursors / Q000378:metabolism*; D051381:Rats; D020798:Two-Hybrid System Techniques
D011498:Protein Precursors; D005941:Glucokinase; D019735:Carbon-Carbon Ligases; C022458:propionyl CoA carboxylase (ATP-hydrolyzing)