Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Sep;87(17):6781–6785. doi: 10.1073/pnas.87.17.6781

Processing of prosecretin: isolation of a secretin precursor from porcine intestine.

G Gafvelin 1, H Jörnvall 1, V Mutt 1
PMCID: PMC54621  PMID: 2395872

Abstract

A precursor to the gastrointestinal hormone secretin has been isolated. The starting material for the purification of the precursor was a peptide fraction purified from pig intestinal extracts, containing peptides with a molecular weight higher than that of secretin. The purification could be followed by measurement of secretin bioactivity (alkali secreted in the pancreatic juice of anesthetized cat). Sequence analysis of the isolated secretin precursor revealed a 71-amino acid residue polypeptide that contained the sequence of secretin N terminally, followed by a Gly-Lys-Arg sequence and a C-terminal extension of 41-amino acid residues. With the exception of an arginine residue, which occurs directly after the Gly-Lys-Arg sequence, the remainder of the C-terminal residues in this precursor are identical to the 40 C-terminal residues predicted by the recently described cDNA sequence for porcine preprosecretin. Compared to the deduced preprosecretin sequence, a stretch of 32 amino acid residues directly following the Gly-Lys-Arg sequence is missing in the now purified secretin precursor. This implies that differential splicing may occur when the secretin gene transcript is processed to mRNA.

Full text

PDF
6784

Selected References

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

  1. Adelson J. W., Nelbach L., Yates G. B., Ehrlich A., Glaser C. B., Chang R. Purification and characterization of chymodenin. A hormone-like peptide from porcine duodenum. J Biol Chem. 1986 Aug 15;261(23):10569–10575. [PubMed] [Google Scholar]
  2. Amara S. G., Jonas V., Rosenfeld M. G., Ong E. S., Evans R. M. Alternative RNA processing in calcitonin gene expression generates mRNAs encoding different polypeptide products. Nature. 1982 Jul 15;298(5871):240–244. doi: 10.1038/298240a0. [DOI] [PubMed] [Google Scholar]
  3. Bayliss W. M., Starling E. H. The mechanism of pancreatic secretion. J Physiol. 1902 Sep 12;28(5):325–353. doi: 10.1113/jphysiol.1902.sp000920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bell G. I., Santerre R. F., Mullenbach G. T. Hamster preproglucagon contains the sequence of glucagon and two related peptides. Nature. 1983 Apr 21;302(5910):716–718. doi: 10.1038/302716a0. [DOI] [PubMed] [Google Scholar]
  5. Bodner M., Fridkin M., Gozes I. Coding sequences for vasoactive intestinal peptide and PHM-27 peptide are located on two adjacent exons in the human genome. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3548–3551. doi: 10.1073/pnas.82.11.3548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bradbury A. F., Smyth D. G. Enzyme-catalysed peptide amidation. Isolation of a stable intermediate formed by reaction of the amidating enzyme with an imino acid. Eur J Biochem. 1987 Dec 15;169(3):579–584. doi: 10.1111/j.1432-1033.1987.tb13648.x. [DOI] [PubMed] [Google Scholar]
  7. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  8. Buscail L., Cauvin A., Gourlet P., Gossen D., De Neef P., Rathé J., Robberecht P., Vandermeers-Piret M. C., Vandermeers A., Christophe J. Purification and amino acid sequence of vasoactive intestinal peptide, peptide histidine isoleucinamide (1-27) and secretin from the small intestine of guinea pig. Biochim Biophys Acta. 1990 May 8;1038(3):355–359. doi: 10.1016/0167-4838(90)90248-e. [DOI] [PubMed] [Google Scholar]
  9. Carlquist M., Jörnvall H., Mutt V. Isolation and amino acid sequence of bovine secretin. FEBS Lett. 1981 May 5;127(1):71–74. doi: 10.1016/0014-5793(81)80343-2. [DOI] [PubMed] [Google Scholar]
  10. Carlquist M., Rökaeus A. Isolation of a proform of porcine secretin by ion-exchange and reversed-phase high-performance liquid chromatography. J Chromatogr. 1984 Jul 27;296:143–151. doi: 10.1016/s0021-9673(01)96408-8. [DOI] [PubMed] [Google Scholar]
  11. Chang T. M., Berger-Ornstein L., Chey W. Y. Presence of biologically and immunologically active secretin-like substance in the mammalian brain. Peptides. 1985 Mar-Apr;6(2):193–198. doi: 10.1016/0196-9781(85)90039-7. [DOI] [PubMed] [Google Scholar]
  12. Eipper B. A., Park L. P., Dickerson I. M., Keutmann H. T., Thiele E. A., Rodriguez H., Schofield P. R., Mains R. E. Structure of the precursor to an enzyme mediating COOH-terminal amidation in peptide biosynthesis. Mol Endocrinol. 1987 Nov;1(11):777–790. doi: 10.1210/mend-1-11-777. [DOI] [PubMed] [Google Scholar]
  13. Gafvelin G., Carlquist M., Mutt V. A proform of secretin with high secretin-like bioactivity. FEBS Lett. 1985 May 20;184(2):347–352. doi: 10.1016/0014-5793(85)80636-0. [DOI] [PubMed] [Google Scholar]
  14. Gossen D., Buscail L., Cauvin A., Gourlet P., De Neef P., Rathe J., Robberecht P., Vandermeers-Piret M. C., Vandermeers A., Christophe J. Amino acid sequence of VIP, PHI and secretin from the rabbit small intestine. Peptides. 1990 Jan-Feb;11(1):123–128. doi: 10.1016/0196-9781(90)90120-t. [DOI] [PubMed] [Google Scholar]
  15. Gossen D., Vandermeers A., Vandermeers-Piret M. C., Rathé J., Cauvin A., Robberecht P., Christophe J. Isolation and primary structure of rat secretin. Biochem Biophys Res Commun. 1989 Apr 28;160(2):862–867. doi: 10.1016/0006-291x(89)92514-x. [DOI] [PubMed] [Google Scholar]
  16. Gubler U., Monahan J. J., Lomedico P. T., Bhatt R. S., Collier K. J., Hoffman B. J., Böhlen P., Esch F., Ling N., Zeytin F. Cloning and sequence analysis of cDNA for the precursor of human growth hormone-releasing factor, somatocrinin. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4311–4314. doi: 10.1073/pnas.80.14.4311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Heinrich G., Gros P., Habener J. F. Glucagon gene sequence. Four of six exons encode separate functional domains of rat pre-proglucagon. J Biol Chem. 1984 Nov 25;259(22):14082–14087. [PubMed] [Google Scholar]
  18. Itoh N., Obata K., Yanaihara N., Okamoto H. Human preprovasoactive intestinal polypeptide contains a novel PHI-27-like peptide, PHM-27. Nature. 1983 Aug 11;304(5926):547–549. doi: 10.1038/304547a0. [DOI] [PubMed] [Google Scholar]
  19. Kaiser R., Holmquist B., Hempel J., Vallee B. L., Jörnvall H. Class III human liver alcohol dehydrogenase: a novel structural type equidistantly related to the class I and class II enzymes. Biochemistry. 1988 Feb 23;27(4):1132–1140. doi: 10.1021/bi00404a009. [DOI] [PubMed] [Google Scholar]
  20. Kawaguchi Y., Hoshimaru M., Nawa H., Nakanishi S. Sequence analysis of cloned cDNA for rat substance P precursor: existence of a third substance P precursor. Biochem Biophys Res Commun. 1986 Sep 30;139(3):1040–1046. doi: 10.1016/s0006-291x(86)80282-0. [DOI] [PubMed] [Google Scholar]
  21. Kimura C., Ohkubo S., Ogi K., Hosoya M., Itoh Y., Onda H., Miyata A., Jiang L., Dahl R. R., Stibbs H. H. A novel peptide which stimulates adenylate cyclase: molecular cloning and characterization of the ovine and human cDNAs. Biochem Biophys Res Commun. 1990 Jan 15;166(1):81–89. doi: 10.1016/0006-291x(90)91914-e. [DOI] [PubMed] [Google Scholar]
  22. Kitamura N., Kitagawa H., Fukushima D., Takagaki Y., Miyata T., Nakanishi S. Structural organization of the human kininogen gene and a model for its evolution. J Biol Chem. 1985 Jul 15;260(14):8610–8617. [PubMed] [Google Scholar]
  23. Kopin A. S., Wheeler M. B., Leiter A. B. Secretin: structure of the precursor and tissue distribution of the mRNA. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2299–2303. doi: 10.1073/pnas.87.6.2299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Linder S., Barkhem T., Norberg A., Persson H., Schalling M., Hökfelt T., Magnusson G. Structure and expression of the gene encoding the vasoactive intestinal peptide precursor. Proc Natl Acad Sci U S A. 1987 Jan;84(2):605–609. doi: 10.1073/pnas.84.2.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lopez L. C., Frazier M. L., Su C. J., Kumar A., Saunders G. F. Mammalian pancreatic preproglucagon contains three glucagon-related peptides. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5485–5489. doi: 10.1073/pnas.80.18.5485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mayo K. E., Vale W., Rivier J., Rosenfeld M. G., Evans R. M. Expression-cloning and sequence of a cDNA encoding human growth hormone-releasing factor. Nature. 1983 Nov 3;306(5938):86–88. doi: 10.1038/306086a0. [DOI] [PubMed] [Google Scholar]
  27. Mutt V., Carlquist M., Tatemoto K. Secretin-like bioactivity in extracts of porcine brain. Life Sci. 1979 Nov 12;25(20):1703–1707. doi: 10.1016/0024-3205(79)90472-7. [DOI] [PubMed] [Google Scholar]
  28. Mutt V., Jorpes J. E., Magnusson S. Structure of porcine secretin. The amino acid sequence. Eur J Biochem. 1970 Sep;15(3):513–519. doi: 10.1111/j.1432-1033.1970.tb01034.x. [DOI] [PubMed] [Google Scholar]
  29. Nawa H., Hirose T., Takashima H., Inayama S., Nakanishi S. Nucleotide sequences of cloned cDNAs for two types of bovine brain substance P precursor. Nature. 1983 Nov 3;306(5938):32–36. doi: 10.1038/306032a0. [DOI] [PubMed] [Google Scholar]
  30. Nilsson A., Carlquist M., Jörnvall H., Mutt V. Isolation and characterization of chicken secretin. Eur J Biochem. 1980 Nov;112(2):383–388. doi: 10.1111/j.1432-1033.1980.tb07216.x. [DOI] [PubMed] [Google Scholar]
  31. O'Donohue T. L., Charlton C. G., Miller R. L., Boden G., Jacobowitz D. M. Identification, characterization, and distribution of secretin immunoreactivity in rat and pig brain. Proc Natl Acad Sci U S A. 1981 Aug;78(8):5221–5224. doi: 10.1073/pnas.78.8.5221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sausville E. A., Lebacq-Verheyden A. M., Spindel E. R., Cuttitta F., Gazdar A. F., Battey J. F. Expression of the gastrin-releasing peptide gene in human small cell lung cancer. Evidence for alternative processing resulting in three distinct mRNAs. J Biol Chem. 1986 Feb 15;261(5):2451–2457. [PubMed] [Google Scholar]
  33. Shinomura Y., Eng J., Yalow R. S. Dog secretin: sequence and biologic activity. Life Sci. 1987 Sep 7;41(10):1243–1248. doi: 10.1016/0024-3205(87)90202-5. [DOI] [PubMed] [Google Scholar]
  34. Spindel E. R., Zilberberg M. D., Habener J. F., Chin W. W. Two prohormones for gastrin-releasing peptide are encoded by two mRNAs differing by 19 nucleotides. Proc Natl Acad Sci U S A. 1986 Jan;83(1):19–23. doi: 10.1073/pnas.83.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Straus E., Yalow R. S. Immunoreactive secretin in gastrointestinal mucosa of several mammalian species. Gastroenterology. 1978 Sep;75(3):401–404. [PubMed] [Google Scholar]
  36. Takeda J., Seino Y., Tanaka K., Fukumoto H., Kayano T., Takahashi H., Mitani T., Kurono M., Suzuki T., Tobe T. Sequence of an intestinal cDNA encoding human gastric inhibitory polypeptide precursor. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7005–7008. doi: 10.1073/pnas.84.20.7005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. White J. W., Saunders G. F. Structure of the human glucagon gene. Nucleic Acids Res. 1986 Jun 25;14(12):4719–4730. doi: 10.1093/nar/14.12.4719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. van den Eijnden-van Raaij A. J., Koornneef I., van Oostwaard T. M., de Laat S. W., van Zoelen E. J. Cation-exchange high-performance liquid chromatography: separation of highly basic proteins using volatile acidic solvents. Anal Biochem. 1987 May 15;163(1):263–269. doi: 10.1016/0003-2697(87)90122-9. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES