Summary
The insulin storage granule of the pancreatic beta cell is assembled within the trans Golgi network from around 50 or so gene products many of which are synthesized coordinately with the major component, proinsulin. An important contribution to our understanding of the regulation of this process has come from studies of the post-translational processing of proinsulin and of other proteins which are stored in the granule, particularly the processing enzymes themselves. The present review focusses on recent insights into the molecular nature of the processing machinery, and the granule Ca2+-dependent subtilisin-related endopeptidases which catalyse the initial rate-limiting step in the enzymic conversion of proinsulin.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- TGN:
-
Trans Golgi network
- CPH:
-
carboxypeptidase H
- NMR:
-
nuclear magnetic resonance
- Lys:
-
lysine
- Arg:
-
arginine, Thr, threonine
- Leu:
-
leucine
- Gln:
-
glycine
- Asp:
-
asparginine
- His:
-
histidine
- Ser:
-
serine
- POMC:
-
pro-opiomelanocortin
References
Burgess TL, Kelly RB (1987) Constitutive and regulated secretion of proteins. Ann Rev Cell Biol 3: 243–294
De Camilli P, Jahn R (1990) Pathways to regulated exocytosis in neurons. Ann Rev Physiol 52: 625–645
Griffiths G, Simons K (1986) The trans Golgi network: sorting at the exit site of the Golgi complex. Science 234: 438–443
Shields D (1991) Signals for intracellular protein sorting: implications for peptide hormone precursor processing and secretion. In: Fricker LD (ed) Peptide biosynthesis and processing CRC Press, Boca Raton, London pp 37–70
Orci L, Ravazzola M, Amherdt M, Madsen O, Vassalli JD, Perrelet A (1985) Direct identification of prohormone conversion site in insulin-secreting cells. Cell 42: 671–681
Orci L, Ravazzola M, Amherdt M, Madsen O, Perrelet A, Vassalli JD, Anderson RG (1986) Conversion of proinsulin to insulin occurs co-ordinately with acidification of maturing secretory granules. J Cell Biol 103: 2273–2281
Hutton JC (1982) The internal pH and membrane potential of the insulin secretory granule. Biochem J 20: 171–178
Arvan P, Castle D (1992) Protein sorting and secretion granule formation in regulated secretory cells. Trends Cell Biol 2: 327–331
von Zastrow M, Castle JD (1987) Protein sorting among two distinct export pathways occurs from the content of maturing exocrine storage granules. J Cell Biol 105: 2675–2684
Stoller TJ, Shields D (1989) Retrovirus-mediated expression of preprosomatostatin: posttranslational processing, intracellular storage and secretion in GH3 cells. J Cell Biol 107: 2087–2095
Wagner DD, Saffaripour S, Bonfanti R, Sadler JE, Cramer EM, Chapman B, Mayadas TN (1991) Induction of specific storage organelles by von Willebrand factor propolypeptide. Cell 64: 403–413
Powell SK, Orci L, Craik CS, Moore H-PH (1988) Efficient targeting to storage granules of human proinsulins with altered propeptide domain. J Cell Biol 106: 1843–1851
Shakur Y, Shennan KI, Taylor NA, Docherty K (1989) A major C-peptide deletion prevents secretion of a mutant human proinsulin from transfected monkey kidney cells. J Mol Endocrinol 3: 155–162
Gross DJ, Villa-Komaroff L, Kahn CR, Weir GC, Halban PA (1989) Deletion of a highly conserved tetrapeptide sequence of the proinsulin connecting peptide (C-peptide) inhibits proinsulin to insulin conversion by transfected pituitary corticotroph (AtT20) cells. J Biol Chem 264: 21486–21490
Gross DJ, Halban PA, Kahn CR, Weir GC, Villa Komaroff L (1989) Partial diversion of a mutant proinsulin (B10 aspartic acid) from the regulated to the constitutive secretory pathway in transfected AtT-20 cells. Proc Natl Acad Sci USA 86: 4107–4111
Orci L, Ravazzola M, Perrelet A (1984) Proinsulin associates with Golgi membranes of the pancreatic β-cell. Proc Natl Acad Sci USA 81: 6743–6746
Chung K-N, Walter P, Aponte GW, Moore H-P (1989) Molecular sorting in the secretory pathway. Science 243: 192–197
Rosa P, Weiss U, Pepperkok R, Ansorge W, Niehrs C, Stelzer EHK, Huttner WB (1989) An antibody against secretogranin 1 (chromogranin B) is packaged into secretory granules. J Cell Biol 109: 17–34
Hutton JC (1989) The 24th Minkowski lecture: The insulin secretory granule Diabetologia 32: 271–281
Hutton JC (1984) Secretory granules Experientia 40: 1091–1098
Guest PC, Rhodes CJ, Hutton JC (1989) Regulation of the biosynthesis of insulin secretory granule proteins: co-ordinate translational control is exerted on some, but not all, granule matrix constituents. Biochem J 257: 431–437
Guest PC, Bailyes EM, Rutherford NG, Hutton JC (1990) Insulin secretory granule biogenesis: co-ordinate regulation of the biosynthesis of the majority of constituent proteins. Biochem J 274: 73–78
Sopwith AM, Hales CN, Hutton JC (1984) Pancreatic B-cells secrete a range of novel proteins besides insulin. Biochim Biophys Acta 803: 342–345
Guest PC, Pipeleers D, Rossier J, Rhodes CJ, Hutton JC (1990) Co-secretion of carboxypeptidase H and insulin from isolated rat islets of Langerhans. Biochem J 264: 503–508
Fricker LD (1991) Peptide processing exopeptidases: amino and carboxypeptidases involved with peptide biosynthesis. In: Fricker LD (ed) Peptide biosynthesis and processing. CRC Press, Boca Raton, London pp 199–229
Winkler H, Hortnagl H, Smith AD (1970) Membranes of the adrenal medulla. Biochem J 118: 303–310
Hunter A, Phillips JH (1989) The recycling of a secretory granule membrane protein. Exp Cell Res 182: 445–460
Brand SH, Laurie SM, Mixon MB, Castle JD (1991) Secretory carrier membrane proteins 31–35 define a common protein composition among secretory carrier membranes. J Biol Chem 266: 18 949–18 957
Rudnick G (1986) ATP-driven H+ pumping into intracellular organelles. Ann Rev Physiol 48: 403–413
Grimaldi KA, Hutton JC, Siddle K (1987) Production and characterisation of monoclonal antibodies to insulin secretory granule membrane. Biochem J 245: 567–573
Grimaldi KA, Siddle K, Hutton JC (1987) Biosynthesis of insulin secretory granule membrane proteins: control by glucose. Biochem J 245: 557–566
Davidson HW, Rhodes CJ, Hutton JC (1988) Intraorganellar calcium and pH control proinsulin cleavage in the pancreatic β-cell via two distinct site-specific endopeptidases. Nature (London) 333: 93–96
Rhodes CJ, Zumbrunn A, Bailyes EM, Shaw E, Hutton JC (1989) The inhibition of proinsulin-processing endopeptidase activities by active-site-directed peptides. Biochem J 258: 305–308
Taylor NA, Docherty K (1992) Sequence requirements for processing of proinsulin in transfected mouse pituitary AtT20 cells. Biochem J 286: 619–622
Rhodes CJ, Lincoln B, Shoelson SE (1992) Preferential cleavage of des-31, 32-proinsulin over intact proinsulin by the insulin secretory granule type 2 endopeptidase: implications of a favoured route for prohormone processing. J Biol Chem 267: 22 719–22 727
Mizuno K, Nakamura T, Oshima T, Tanaka S, Matsuo H (1988) Yeast KEX2 gene encodes an endopeptidase homologous to subtilisin like serine proteases. Biochem Biophys Res Commun 156: 246–254
Roebroek AJM, Schalken JA, Bussemakers MJG et al. (1986) Characterisation of human c-fes/fps reveals a new transcription unit (fur) in the immediately upstream region of the proto-oncogene. Mol Biol Rep 11: 117–125
Molloy SS, Bresnahan PA, Leppla SH, Klimpel KR, Thomas G (1992) Human furin is a calcium-dependent serine endoprotease that recognises the sequence Arg-X-X-Arg and efficiently cleaves anthrax toxin protective antigen. J Biol Chem 267: 16 396–16 402
Wise RJ, Barr PJ, Wong PA, Kiefer MC, Brake AJ, Kaufman RJ (1990) Expression of human proprotein processing enzyme: correct cleavage of the von Willebrand factor precursor at a paired basic amino acid site. Proc Natl Acad Sci USA 87: 9378–9382
Bresnahan PA, Leduc R, Thomas L et al. (1990) Human fur gene encodes a yeast KEX-2-like endoprotease that cleaves pro-β-NGF in vivo. J Cell Biol 111: 2851–2859
Misumi Y, Oda K, Fujiwara T, Takami N, Tashiro K, Ikehara Y (1991) Functional expression of furin demonstrating its intracellular localisation and endoprotease activity for processing of proalbumin and complement pro-C3. J Biol Chem 266: 16 954–16 959
Smeekens SP, Avruch AS, La Mendola J, Chan SJ, Steiner DF (1991) Identification of a cDNA encoding a putative prohormone convertase related to PC2 in AtT20 cells and islets of Langerhans. Proc Natl Acad Sci USA 88: 340–344
Seidah NG, Gaspar L, Mion P, Marcinkiewicz M, Mbikay M, Chretien M (1990) cDNA sequence of two distinct pituitary proteins homologous to Kex2 and furin gene products: tissue-specific mRNAs encoding candidates for pro-hormone processing proteinases. DNA Cell Biol 9: 415–424
Fricker LD, Das B, Hogue-Angeletti R (1990) Identification of the pH-dependent membrane anchor of carboxypeptidase E (EC 3.4.17.10). J Biol Chem 265: 2476–2482
Thomas L, Leduc R, Thorne BA, Smeekens SP, Steiner DF, Thomas G (1991) Kex2-like endoproteases PC2 and PC3 accurately cleave a model prohormone in mammalian cells: Evidence for a common core of neuroendocrine processing enzymes. Proc Natl Acad Sci USA 88: 5297–5301
Nakayama K, Hosaka M, Hatsuzawa K, Murakami K (1991) Cloning and functional expression of a novel endoprotease involved in prohormone processing at dibasic sites. J Biochem 109: 803–806
Nakayama K, Watanabe T, Nakagawa et al. (1992) Consensus sequence for precursor processing at mono-arginyl sites: evidence for the involvement of a Kex2-like endoprotease in precursor cleavages at both dibasic and mono-arginyl sites. J Biol Chem 267: 16 335–16 340
Shennan KIJ, Smeekens SP, Steiner DF, Docherty K (1991) Characterisation of PC2, a mammalian Kex2 homologue, following expression of the cDNA in microinjected Xenopus oocytes. FEBS Lett 284: 277–280
Bailyes EM, Shennan KIJ, Seal AJ et al. (1992) A member of the eukaryotic subtilisin family has the enzymatic properties of the type 1 proinsulin-converting endopeptidase. Biochem J 285: 391–394
Bennett DL, Bailyes EM, Nielsen E, Guest PC, Rutherford NG, Arden SD, Hutton JC (1992) Identification of the type 2 proinsulin processing endopeptidase as PC2, a member of the eukaryote subtilisin family. J Biol Chem 267: 15229–15236
Smeekens SP, Montag AG, Thomas G et al. (1992) Proinsulin processing by the subtilisin-related proprotein convertases furin, PC2 and PC3. Proc Natl Acad Sci USA 89: 8822–8826
Nakagawa T, Hosaka M, Torii S, Watanabe T, Murakami K, Nakagawa K (1993) Identification and functional expression of a new member of the mammalian Kex2-like processing endoprotease family: its striking structural similarity to PACE4. J Biochem 113: 132–135
Seidah NG, Day R, Hamelin J, Gaspar A, Collard MW, Chretien M (1992) Testicular expression of PC4 in the rat: molecular diversity of a novel germ cell-specific Kex2/subtilisin-like proprotein convertase. Mol Endocrinol 6: 1559–1570
Nakayama K, Kim W-S, Torii S et al. (1992) Identification of the fourth member of the mammalian endoprotease family homologous to the yeast Kex2 protease: its testisspecific expression. J Biol Chem 267: 5897–5900
Seidah NG, Day R, Marcinkiewicz M, Benjannet S, Chretien M (1991) Mammalian neural and endocrine pro-protein and pro-hormone convertases belonging to the subtilisin family of serine proteinases. Enzyme 45: 271–284
Christie DL, Batchelor DC, Palmer DJ (1991) Identification of Kex-2-related proteases in chromaffin granules by partial amino acid sequence analysis. J Biol Chem 266: 15 679–15 683
Mackin RB, Noe BD, Speiss J (1991) Identification of a somatostatin-14-generating propeptide converting enzyme as a member of the Kex2/furin/PC family. Endocrinology (Baltimore) 129: 2263–2265
Guest PC, Arden SD, Bennett DL, Clark A, Rutherford NG, Hutton JC (1992) The post-translational processing and intracellular sorting of PC2 in the islets of Langerhans. J Biol Chem 267: 22401–22406
Shennan KIJ, Seal AJ, Smeekens SP, Steiner DF, Docherty K (1991) Site-directed mutagenesis and expression of PC2 in microinjected Xenopus oocytes. J Biol Chem 266: 24 011–24 017
Vindrola O, Lindberg I (1992) Biosynthesis of the prohormone convertase mPC1 in AtT20 cells. Mol Endocrinol 6: 1088–1094
Steiner DF, Smeekens SP, Ohagi S, Chan SJ (1992) The new enzymology of precursor processing endoproteases. J Biol Chem 267: 23 435–23 438
Alarcon C, Lincoln B, Rhodes CJ (1993) The biosynthesis of the subtilisin-related proprotein convertase PC3, but not that of the PC2 convertase, is regulated by glucose in parallel to proinsulin biosynthesis in rat pancreatic islets. J Biol Chem 268: 4276–4280
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hutton, J.C. Insulin secretory granule biogenesis and the proinsulin-processing endopeptidases. Diabetologia 37 (Suppl 2), S48–S56 (1994). https://doi.org/10.1007/BF00400826
Issue Date:
DOI: https://doi.org/10.1007/BF00400826