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. 1986 Mar 15;234(3):665–670. doi: 10.1042/bj2340665

Trypsin-catalysed formation of pig des-(23-63)-proinsulin from desoctapeptide-(B23-30)-insulin.

T Kubiak, D Cowburn
PMCID: PMC1146623  PMID: 3521594

Abstract

Incubation of pig desoctapeptide-(B23-30)-insulin with trypsin in solvent systems consisting of dimethyl sulphoxide, butane-1,4-diol and Tris buffer resulted in the formation of an extra peptide bond between Arg-B22 and Gly-A1 in the DOPI molecule. This DOPI derivative can also be regarded as pig des-(23-63)-proinsulin. The structure of the new, previously unreported, proinsulin analogue was determined on the basis of amino acid analysis, dansylation and digestion with Staphylococcus aureus V8 proteinase. Receptor-binding ability of des-(23-63)-proinsulin was 20% of that of pig desoctapeptide-(B23-30)-insulin and 0.02% of that of pig insulin.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cao Q. P., Cui D. F., Zhang Y. S. Enzymatic synthesis of deshexapeptide insulin. Nature. 1981 Aug 20;292(5825):774–775. doi: 10.1038/292774a0. [DOI] [PubMed] [Google Scholar]
  2. Chance R. E., Ellis R. M., Bromer W. W. Porcine proinsulin: characterization and amino acid sequence. Science. 1968 Jul 12;161(3837):165–167. doi: 10.1126/science.161.3837.165. [DOI] [PubMed] [Google Scholar]
  3. Cui D. F., Cao Q. P., Zhu S. Q., Zhang X. T., Zhang Y. S. Enzymatic synthesis of deshexapeptide insulin and its analogues. Sci Sin B. 1983 Mar;26(3):248–254. [PubMed] [Google Scholar]
  4. Gattner H. G., Danho W., Behn C., Zahn H. The preparation of two mutant forms of human insulin, containing leucine in position B24 or B25, by enzyme-assisted synthesis. Hoppe Seylers Z Physiol Chem. 1980 Jul;361(7):1135–1138. [PubMed] [Google Scholar]
  5. Houmard J., Drapeau G. R. Staphylococcal protease: a proteolytic enzyme specific for glutamoyl bonds. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3506–3509. doi: 10.1073/pnas.69.12.3506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Inouye K., Watanabe K., Tochino Y., Kanaya T., Kobayashi M., Shigeta Y. Semisynthesis and biological properties of the [B24-leucine]-, [B25-leucine[- and [B24-leucine, B25-leucine]-analogues of human insulin. Experientia. 1981;37(8):811–813. doi: 10.1007/BF01985653. [DOI] [PubMed] [Google Scholar]
  7. Inouye K., Watanabe K., Tochino Y., Kobayashi M., Shigeta Y. Semisynthesis and properties of some insulin analogs. Biopolymers. 1981 Sep;20(9):1845–1858. doi: 10.1002/bip.1981.360200909. [DOI] [PubMed] [Google Scholar]
  8. Kobayashi M., Haneda M., Maegawa H., Watanabe N., Takada Y., Shigeta Y., Inouye K. Receptor binding and biological activity of [SerB24]-insulin, an abnormal mutant insulin. Biochem Biophys Res Commun. 1984 Feb 29;119(1):49–57. doi: 10.1016/0006-291x(84)91616-4. [DOI] [PubMed] [Google Scholar]
  9. Markussen J., Jørgensen K. H., Sørensen A. R., Thim L. Single chain des-(B30) insulin. Intramolecular crosslinking of insulin by trypsin catalyzed transpeptidation. Int J Pept Protein Res. 1985 Jul;26(1):70–77. [PubMed] [Google Scholar]
  10. McCaleb M. L., Donner D. B. Affinity of the hepatic insulin receptor is influenced by membrane phospholipids. J Biol Chem. 1981 Nov 10;256(21):11051–11057. [PubMed] [Google Scholar]
  11. Shaw W. N., Chance R. E. Effect of porcine proinsulin in vitro on adipose tissue and diaphragm of the normal rat. Diabetes. 1968 Dec;17(12):737–745. doi: 10.2337/diab.17.12.737. [DOI] [PubMed] [Google Scholar]
  12. Shoelson S., Fickova M., Haneda M., Nahum A., Musso G., Kaiser E. T., Rubenstein A. H., Tager H. Identification of a mutant human insulin predicted to contain a serine-for-phenylalanine substitution. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7390–7394. doi: 10.1073/pnas.80.24.7390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Tager H., Thomas N., Assoian R., Rubenstein A., Saekow M., Olefsky J., Kaiser E. T. Semisynthesis and biological activity of porcine [LeuB24]insulin and [LeuB25]insulin. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3181–3185. doi: 10.1073/pnas.77.6.3181. [DOI] [PMC free article] [PubMed] [Google Scholar]

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