Severe defect in proglucagon processing in islet A-cells of prohormone convertase 2 null mice.
Mice homozygous for a deletion in the gene encoding prohormone convertase 2 (PC2) are generally healthy but have mild hypoglycemia and flat glucose-tolerance curves. Their islets show marked alpha (A)-cell hyperplasia, suggesting a possible defect in glucagon processing (Furuta, M., Yano, H., Zhou, A., Rouille, Y., Holst, J., Carroll, R., Ravazzola, M., Orci, L., Furuta, H., and Steiner, D. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 6646-6651). In this report we have examined the biosynthesis and processing of proglucagon in isolated islets from these mice via pulse-chase labeling and find that proglucagon undergoes essentially no processing in chase periods up to 8 h in duration. Only a small percent of cleavage at the sensitive interdomain site (residues 71 and 72) appears to occur. These observations thus conclusively demonstrate the essentiality of PC2 for the production of glucagon in the islet A-cells. Ultrastructural and immunocytochemical studies indicate the presence of large amounts of proglucagon in atypical appearing secretory granules in the hyperplastic and hypertrophic A-cells, along with morphological evidence of high rates of proglucagon secretion in PC2 null islets. These findings provide strong evidence that active glucagon is required to maintain normal blood glucose levels, counterbalancing the action of insulin at all times.
Furuta
M
M
Department of Biochemistry and Molecular Biology and the Howard Hughes Medical Institute, University of Chicago, Chicago, Illinois 60637, USA.
Zhou
A
A
Webb
G
G
Carroll
R
R
Ravazzola
M
M
Orci
L
L
Steiner
D F
DF
D000818:Animals; D015153:Blotting, Western; D005934:Glucagon / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D007515:Islets of Langerhans / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D051379:Mice; D008817:Mice, Mutant Strains; D052336:Proglucagon; D045706:Proprotein Convertase 2; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D013381:Subtilisins / Q000378:metabolism*
D011498:Protein Precursors; D012333:RNA, Messenger; D052336:Proglucagon; D005934:Glucagon; D013381:Subtilisins; D045706:Proprotein Convertase 2