Regulation of the glutamate dehydrogenase activity in rat islets of Langerhans and its consequence on insulin release.
Kinetic properties of glutamate dehydrogenase (GDH) and the effects on its activity of several putative modulators were examined in mitochondrial extracts of rat pancreatic islets. In the presence of 40 mmol/L NH4Cl and 0.1 mmol/L NADH, stepwise elevation of the 2-oxoglutarate concentration from 0.005 to 0.05 mmol/L increased glutamate formation, whereas further increases led to a progressive decrease of the reaction velocity. Adenosine diphosphate (ADP) at 0.1 mmol/L partially and at 1 mmol/L completely reversed the inhibitory effect of 2-oxoglutarate. The sensitivity to activation by either ADP or leucine was dependent on 2-oxoglutarate concentrations. At higher concentrations of the latter, greater amounts of the activators were needed to attain maximal effect. In the absence of allosteric activators, sulfate or phosphate at 20 mmol/L partially released the inhibitory effect of 2-oxoglutarate levels and increased the maximal velocity (Vmax) for the reaction. In the presence of 0.1 mmol/L ADP, both anions prevented the inhibition by higher concentrations of 2-oxoglutarate, whereas with 1 mmol/L ADP their only effect was a slight increase in the Vmax. Mg2+ and naturally occurring polyamines decreased glutamate formation in a dose-dependent manner; with 0.1 mmol/L ADP, inhibition was seen at all 2-oxoglutarate concentrations studied, whereas with 1 mmol/L ADP, it was noticeable at substrate concentrations higher than 0.5 mmol/L. This inhibitory effect on GDH activity was partially attenuated by sulfate. Addition of either 2 mmol/L spermidine or extra magnesium (final 2.5 or 5 mmol/L) to the perifusion buffer markedly attenuated the insulin release elicited by alpha-ketoisocaproate. It is suggested that naturally occurring polyamines, magnesium, and phosphate act as physiological modulators of GDH activity in pancreatic beta cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Bryła
J
J
Department of Pharmacology, University of Pennsylvania, School of Medicine, Philadelphia 19104-6084.
Michalik
M
M
Nelson
J
J
Erecińska
M
M
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D005969:Glutamate Dehydrogenase / Q000378:metabolism*; D005971:Glutamates / Q000096:biosynthesis; D018698:Glutamic Acid; D007328:Insulin / Q000378:metabolism*; D007515:Islets of Langerhans / Q000378:metabolism*; D007656:Ketoglutaric Acids / Q000378:metabolism; D007700:Kinetics; D007930:Leucine / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D008297:Male; D008928:Mitochondria / Q000378:metabolism; D009994:Osmolar Concentration; D011073:Polyamines / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013347:Subcellular Fractions / Q000378:metabolism
D005971:Glutamates; D007328:Insulin; D007656:Ketoglutaric Acids; D011073:Polyamines; D018698:Glutamic Acid; D000244:Adenosine Diphosphate; D005969:Glutamate Dehydrogenase; D007930:Leucine; D008274:Magnesium