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PMID: 2657740 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Partial diversion of a mutant proinsulin (B10 aspartic acid) from the regulated to the constitutive secretory pathway in transfected AtT-20 cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 11 ·1989-06-00 ·Pages 4107-11

Gross DJ, Halban PA, Kahn CR, Weir GC, Villa-Komaroff L

Abstract

A patient with type II diabetes associated with hyperproinsulinemia has been shown to have a point mutation in one insulin gene allele, resulting in replacement of histidine with aspartic acid at position 10 of the B-chain. To investigate the basis of the proinsulin processing defect, we introduced an identical mutation in the rat insulin II gene and expressed both the normal and the mutant genes in the AtT-20 pituitary corticotroph cell line. Cells expressing the mutant gene showed increased secretion of proinsulin relative to insulin and rapid release of newly synthesized proinsulin. Moreover, the mutant cell lines did not store the prohormone nor did they release it upon stimulation with secretagogues. These data indicate that a significant fraction of the mutant prohormone is released via the constitutive secretory pathway rather than the regulated pathway, thereby bypassing granule-related processing and regulated release.

MeSH Terms
Animals Aspartic Acid Cell Line Genes Genetic Vectors Insulin/metabolism Insulin Secretion Mutation Pituitary Neoplasms Plasmids Proinsulin/biosynthesis,genetics,metabolism Rats Transfection
Chemicals
Insulin Aspartic Acid Proinsulin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gross D J
E.P. Joslin Research Laboratory, Joslin Diabetes Center, Boston, MA.
Halban P A
Kahn C R
Weir G C
Villa-Komaroff L
References (30)
30 references, click to expand
  1. High-performance liquid chromatography (HPLC): a rapid, flexible and sensitive method for separating islet proinsulin and insulin.
    Diabetologia. 1986 Dec;29(12):893-6 PMID: 3552815
  2. Glucose stimulates the biosynthesis of rat I and II insulin to an equal extent in isolated pancreatic islets.
    FEBS Lett. 1987 May 4;215(1):179-82 PMID: 3552733
  3. Resistance of the insulin crystal to lysosomal proteases: implications for pancreatic B-cell crinophagy.
    Diabetologia. 1987 May;30(5):348-53 PMID: 3301485
  4. Newly synthesized proinsulin/insulin and stored insulin are released from pancreatic B cells predominantly via a regulated, rather than a constitutive, pathway.
    J Cell Biol. 1987 Jul;105(1):145-53 PMID: 3301864
  5. A superactive insulin: [B10-aspartic acid]insulin(human).
    Proc Natl Acad Sci U S A. 1987 Sep;84(18):6408-11 PMID: 3306677
  6. Sorting of progeny coronavirus from condensed secretory proteins at the exit from the trans-Golgi network of AtT20 cells.
    J Cell Biol. 1987 Sep;105(3):1215-26 PMID: 2821011
  7. High-efficiency transformation of mammalian cells by plasmid DNA.
    Mol Cell Biol. 1987 Aug;7(8):2745-52 PMID: 3670292
  8. The trans-most cisternae of the Golgi complex: a compartment for sorting of secretory and plasma membrane proteins.
    Cell. 1987 Dec 24;51(6):1039-51 PMID: 2826013
  9. A mutant human proinsulin is secreted from islets of Langerhans in increased amounts via an unregulated pathway.
    Proc Natl Acad Sci U S A. 1988 Dec;85(23):8943-7 PMID: 3057496
  10. Oxidation of rat insulin II, but not I, leads to anomalous elution profiles upon HPLC analysis of insulin-related peptides.
    FEBS Lett. 1988 Dec 5;241(1-2):205-8 PMID: 3058512
  11. Interaction of zinc with proinsulin.
    Biochem Biophys Res Commun. 1970 Jan 23;38(2):284-9 PMID: 5418703
  12. A sensitive, precise radioimmunoassay of serum insulin relying on charcoal separation of bound and free hormone moieties.
    Acta Endocrinol (Copenh). 1972 Jul;70(3):487-509 PMID: 5068062
  13. Familial hyperproinsulinemia. An autosomal dominant defect.
    N Engl J Med. 1976 Apr 22;294(17):911-5 PMID: 815812
  14. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  15. Intracellular degradation of insulin stores by rat pancreatic islets in vitro. An alternative pathway for homeostasis of pancreatic insulin content.
    J Biol Chem. 1980 Jul 10;255(13):6003-6 PMID: 6993463
  16. A human proinsulin variant at arginine 65.
    Nature. 1981 Jun 25;291(5817):679-81 PMID: 7242673
  17. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.
    J Mol Appl Genet. 1982;1(4):327-41 PMID: 6286831
  18. Use of recombinant DNA technology to program eukaryotic cells to synthesize rat proinsulin: a rapid expression assay for cloned genes.
    Proc Natl Acad Sci U S A. 1982 Oct;79(19):5798-802 PMID: 6764530
  19. Expressing a human proinsulin cDNA in a mouse ACTH-secreting cell. Intracellular storage, proteolytic processing, and secretion on stimulation.
    Cell. 1983 Dec;35(2 Pt 1):531-8 PMID: 6317196
  20. Insulin, not C-peptide (proinsulin), is present in crinophagic bodies of the pancreatic B-cell.
    J Cell Biol. 1984 Jan;98(1):222-8 PMID: 6368567
  21. Familial hyperproinsulinemia. Two cohorts secreting indistinguishable type II intermediates of proinsulin conversion.
    J Clin Invest. 1984 Mar;73(3):714-9 PMID: 6368587
  22. Familial hyperproinsulinemia due to a proposed defect in conversion of proinsulin to insulin.
    N Engl J Med. 1984 Sep 6;311(10):629-34 PMID: 6382002
  23. Direct identification of prohormone conversion site in insulin-secreting cells.
    Cell. 1985 Sep;42(2):671-81 PMID: 3896518
  24. Pathways of protein secretion in eukaryotes.
    Science. 1985 Oct 4;230(4721):25-32 PMID: 2994224
  25. Re-routing of a secretory protein by fusion with human growth hormone sequences.
    Nature. 1986 May 22-28;321(6068):443-6 PMID: 3012361
  26. Clathrin-coated vesicular transport of secretory proteins during the formation of ACTH-containing secretory granules in AtT20 cells.
    J Cell Biol. 1986 Sep;103(3):839-50 PMID: 3017997
  27. Proinsulin radioimmunoassay in the evaluation of insulinomas and familial hyperproinsulinemia.
    Metabolism. 1986 Dec;35(12):1137-46 PMID: 3023795
  28. Expression of human class II major histocompatibility complex antigens using retrovirus vectors.
    Proc Natl Acad Sci U S A. 1987 Apr;84(8):2150-4 PMID: 3031667
  29. A mutation in the B chain coding region is associated with impaired proinsulin conversion in a family with hyperproinsulinemia.
    Proc Natl Acad Sci U S A. 1987 Apr;84(8):2194-7 PMID: 3470784
  30. Proteolytic maturation of insulin is a post-Golgi event which occurs in acidifying clathrin-coated secretory vesicles.
    Cell. 1987 Jun 19;49(6):865-8 PMID: 3555846
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.11.4107
Published
1989-06-00
Pages
4107-11
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC287398
Subset
IM
Grants
NIDDK NIH HHS · DK35292 · United States
NICHD NIH HHS · PT-HD18655 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.11.4107 PMC Full Text View journal details
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