Insulin secretory granule biogenesis. Co-ordinate regulation of the biosynthesis of the majority of constituent proteins.
Two-dimensional gel-electrophoretic analysis combined with fluorography and densitometric quantification was used to examine the effects of glucose on the biosynthesis of rat pancreatic islet proteins. An increase in the medium glucose concentration from 2.8 to 16.7 mM produced a 10-20 fold stimulation in the synthesis of 10 out of 260 detected islet proteins, as judged by incorporation of [35S]methionine during a 20 min incubation. The synthetic rates of the majority of the remaining proteins were stimulated by 2-4-fold. Greater resolution achieved by pulse-chase labelling and subcellular fractionation showed that, of 32 major proteins localized to insulin secretory granules, the biosynthesis of 25 were stimulated 15-30-fold by glucose. By contrast, only eight of 160 proteins in the soluble fraction showed a response of similar magnitude. It is concluded that there is a major and co-ordinated activation of the biosyntheses of proteins destined for secretory granules, which most likely occurs at the level of translational initiation and signal-recognition-particle-mediated translocation into the endoplasmic reticulum lumen. However, it is clear that not all granule proteins, or the majority of proteins translocated across the endoplasmic reticulum membrane, are affected in an equivalent manner. In addition, the synthesis of a small number of cytosolic proteins may be increased markedly by insulinotropic stimuli.
Guest
P C
PC
Department of Clinical Biochemistry, University of Cambridge, Addenbrooke's Hospital, U.K.
Bailyes
E M
EM
Rutherford
N G
NG
Hutton
J C
JC
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003594:Cytoplasmic Granules / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000096:biosynthesis*; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification; D011865:Radioisotope Dilution Technique; D051381:Rats; D011919:Rats, Inbred Strains; D013462:Sulfur Radioisotopes
D007328:Insulin; D008565:Membrane Proteins; D011506:Proteins; D013462:Sulfur Radioisotopes; D008715:Methionine; D005947:Glucose