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. 1978 Sep;75(9):4115–4119. doi: 10.1073/pnas.75.9.4115

Chemical determination of polypeptide hormones.

K Tatemoto, V Mutt
PMCID: PMC336062  PMID: 279902

Abstract

The presence or absence of peptide hormones in tissue extracts may in certain cases be demonstrated by exposing the extracts to conditions under which characteristic fragments of the polypeptide molecule in question are formed and then analyzing for such fragments. An approximate quantitation of the hormones may also be achieved thereby. In the present work the COOH-terminal fragments of polypeptides containing characteristic alpha-amide groups were released enzymatically and then converted into the fluorescent dansyl derivatives, which were identified by thin-layer chromatography. In this way the presence of secretin, cholecystokinin, and the vasoactive intestinal peptide in concentrates of porcine intestinal extracts were demonstrated by their COOH-terminal amide fragments: valine (or leucylvaline) amide, phenylalanine amide, and asparagine (or leucylasparagine) amide, respectively. The analytical methodology used in the present study may also be useful in devising simple and reliable chemical assay methods for the isolation of already known polypeptides and in the isolation of previously uncharacterized polypeptides from natural sources.

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Selected References

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

  1. Airhart J., Sibiga S., Sanders H., Khairallah E. A. An ultramicro method for quantitation of amino acids in biological fluids. Anal Biochem. 1973 May;53(1):132–140. doi: 10.1016/0003-2697(73)90414-4. [DOI] [PubMed] [Google Scholar]
  2. Brazeau P., Vale W., Burgus R., Ling N., Butcher M., Rivier J., Guillemin R. Hypothalamic polypeptide that inhibits the secretion of immunoreactive pituitary growth hormone. Science. 1973 Jan 5;179(4068):77–79. doi: 10.1126/science.179.4068.77. [DOI] [PubMed] [Google Scholar]
  3. Chang M. M., Leeman S. E., Niall H. D. Amino-acid sequence of substance P. Nat New Biol. 1971 Jul 21;232(29):86–87. doi: 10.1038/newbio232086a0. [DOI] [PubMed] [Google Scholar]
  4. Gruber K. A., Stein S., Brink L., Radhakrishnan A., Udenfriend S. Fluorometric assay of vasopressin and oxytocin: a general approach to the assay of peptides in tissues. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1314–1318. doi: 10.1073/pnas.73.4.1314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jorpes J. E., Mutt V. On the biological assay of secretin. The reference standard. Acta Physiol Scand. 1966 Mar;66(3):316–325. doi: 10.1111/j.1748-1716.1966.tb03205.x. [DOI] [PubMed] [Google Scholar]
  6. Krummen K., Frei R. W. Quantitative analysis of nonapeptides in pharmaceutical dosage forms by high-performance liquid chromatography. J Chromatogr. 1977 Feb 21;132(3):429–436. doi: 10.1016/s0021-9673(00)82907-6. [DOI] [PubMed] [Google Scholar]
  7. Mutt V. Further investigations of intestinal hormonal polypeptides. Clin Endocrinol (Oxf) 1976;5 (Suppl):175S–183S. doi: 10.1111/j.1365-2265.1976.tb03825.x. [DOI] [PubMed] [Google Scholar]
  8. Mutt V., Jorpes J. E. Structure of porcine cholecystokinin-pancreozymin. 1. Cleavage with thrombin and with trypsin. Eur J Biochem. 1968 Oct 17;6(1):156–162. doi: 10.1111/j.1432-1033.1968.tb00433.x. [DOI] [PubMed] [Google Scholar]
  9. Neadle D. J., Pollitt R. J. The formation of 1-dimethylaminonaphthalene-5-sulphonamide during the preparation of 1-dimethylaminonaphthalene-5-sulphonylamino acids. Biochem J. 1965 Nov;97(2):607–608. doi: 10.1042/bj0970607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Parham M. E., Loudon G. M. Carboxyl-terminal sequential degradation of peptides. Biochem Biophys Res Commun. 1978 Jan 13;80(1):1–6. doi: 10.1016/0006-291x(78)91095-1. [DOI] [PubMed] [Google Scholar]
  11. Said S. I., Mutt V. Isolation from porcine-intestinal wall of a vasoactive octacosapeptide related to secretin and to glucagon. Eur J Biochem. 1972 Jul 13;28(2):199–204. doi: 10.1111/j.1432-1033.1972.tb01903.x. [DOI] [PubMed] [Google Scholar]
  12. Walter R. Partial purification and characterization of post-proline cleaving enzyme: enzymatic inactivation of neurohypophyseal hormones by kidney preparations of various species. Biochim Biophys Acta. 1976 Jan 23;422(1):138–158. doi: 10.1016/0005-2744(76)90015-2. [DOI] [PubMed] [Google Scholar]
  13. Woods K. R., Wang K. T. Separation of dansyl-amino acids by polyamide layer chromatography. Biochim Biophys Acta. 1967 Feb 21;133(2):369–370. doi: 10.1016/0005-2795(67)90078-5. [DOI] [PubMed] [Google Scholar]

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