Biosynthesis of insulin secretory granule membrane proteins. Control by glucose.
The biosynthesis of a component SGM 110, specifically localized to the membrane of insulin secretory granules, was studied in rat insulinoma cells and in normal islets of Langerhans. Cells or islets were labelled with [35S]methionine or [3H]mannose and SGM 110 was immunoprecipitated by using a monoclonal antibody. Pulse-chase experiments demonstrated that the nascent polypeptide was cotranslationally glycosylated to form a 97,000 Da peptide which in turn was processed to the mature 110,000 Da form. A 50,000 Da form detected by immunoblotting with the same antibody was not conspicuously labelled even after a 20 h chase incubation, suggesting that it represented late processing of SGM 110 in lysosomes. With insulinoma cells, an increase in medium glucose concentration from 3 mM to 20 mM was without effect on the secretion of insulin or on the biosynthesis of (pro)insulin or SGM 110. In normal islets, however, 20 mM-glucose produced a 17-fold increase in (pro)insulin biosynthesis and a 13-fold increase in SGM 110 biosynthesis, compared with only a 2-fold increase in total protein synthesis, as judged by incorporation of [35S]methionine during a 1 h incubation. The effect of glucose on both (pro)insulin and SGM 110 biosynthesis was blocked by the addition of mannoheptulose, but not by the removal of extracellular calcium, both of which conditions inhibit insulin secretion. In contrast tolbutamide, an agent which stimulates insulin secretion, did not enhance the biosynthesis of (pro)insulin or SGM 110. It is concluded that at least one protein component of the insulin secretory granule membrane is synthesized co-ordinately with proinsulin and is subject to similar regulatory mechanisms. Factors which acutely control insulin secretion may also control granule biogenesis, although the two processes are not coupled in an obligatory fashion.
Grimaldi
K A
KA
Department of Clinical Biochemistry, University of Cambridge, Addenbrooke's Hospital, U.K.
Siddle
K
K
Hutton
J C
JC
D007516:Adenoma, Islet Cell / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D011232:Chemical Precipitation; D003594:Cytoplasmic Granules / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology; D007122:Immunoelectrophoresis; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007340:Insulinoma / Q000378:metabolism*; D008356:Mannoheptulose / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D010190:Pancreatic Neoplasms / Q000378:metabolism*; D011384:Proinsulin / Q000096:biosynthesis; D051381:Rats; D014044:Tolbutamide / Q000494:pharmacology
D007328:Insulin; D008562:Membrane Glycoproteins; C053834:insulin secretory granule membrane glycoprotein 110; D008356:Mannoheptulose; D011384:Proinsulin; D014044:Tolbutamide; D005947:Glucose; D002118:Calcium