Skip to main content
NCBI 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
. 1981 Mar;78(3):1962–1966. doi: 10.1073/pnas.78.3.1962

Two adrenal opioid polypeptides: proposed intermediates in the processing of proenkephalin.

A S Stern, B N Jones, J E Shively, S Stein, S Undenfriend
PMCID: PMC319256  PMID: 6940201

Abstract

Two enkephalin-containing polypeptides of 3600 and 4900 daltons have been isolated from extracts of bovine adrenal medulla, purified to homogeneity, and analyzed by a combination of automated Edman degradation and enzymatic time course hydrolysis. The 4900-dalton polypeptide contains two copies of enkephalin, one an internal [Met]enkephalin sequence, the other a [Leu]enkephalin sequence at the carboxyl terminus. Sequence analysis of the 3600-dalton polypeptide has not been completed, but the polypeptide has been shown to contain a single [Met]enkephalin sequence followed by an -Arg-Phe linkage that forms the carboxyl terminus of the molecule. On the basis of these and other findings, we propose that the above enkephalin-containing polypeptides are intermediates in the biosynthesis of the enkephalins and that they are generated by posttranslational processing from a large multivalent enkephalin precursor molecule, proenkephalin. The term "multivalent" is used to indicate a polypeptide with many identical functional sequences.

Full text

PDF
1962

Selected References

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

  1. Brownstein M. J. Opioid peptides: search for the precursors. Nature. 1980 Oct 23;287(5784):678–679. doi: 10.1038/287678a0. [DOI] [PubMed] [Google Scholar]
  2. Gerber L. D., Stein S., Rubinstein M., Wideman J., Udenfriend S. Binding assay for opioid peptides with neuroblastoma x glioma hybrid cells: specificity of the receptor site. Brain Res. 1978 Jul 28;151(1):117–126. doi: 10.1016/0006-8993(78)90954-x. [DOI] [PubMed] [Google Scholar]
  3. Goldstein A., Tachibana S., Lowney L. I., Hunkapiller M., Hood L. Dynorphin-(1-13), an extraordinarily potent opioid peptide. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6666–6670. doi: 10.1073/pnas.76.12.6666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hayashi R. Carboxypeptidase Y in sequence determination of peptides. Methods Enzymol. 1977;47:84–93. doi: 10.1016/0076-6879(77)47010-1. [DOI] [PubMed] [Google Scholar]
  5. Hughes J., Smith T. W., Kosterlitz H. W., Fothergill L. A., Morgan B. A., Morris H. R. Identification of two related pentapeptides from the brain with potent opiate agonist activity. Nature. 1975 Dec 18;258(5536):577–580. doi: 10.1038/258577a0. [DOI] [PubMed] [Google Scholar]
  6. Hunkapiller M. W., Hood L. E. New protein sequenator with increased sensitivity. Science. 1980 Feb 1;207(4430):523–525. doi: 10.1126/science.7352258. [DOI] [PubMed] [Google Scholar]
  7. Inturrisi C. E., Umans J. G., Wolff D., Stern A. S., Lewis R. V., Stein S., Udenfriend S. Analgesic activity of the naturally occurring heptapeptide [Met]enkephalin-Arg6-Phe7. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5512–5514. doi: 10.1073/pnas.77.9.5512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Johnson N. D., Hunkapiller M. W., Hood L. E. Analysis of phenylthiohydantoin amino acids by high-performance liquid chromatography on DuPont Zobax cyanopropylsilane columns. Anal Biochem. 1979 Dec;100(2):335–338. doi: 10.1016/0003-2697(79)90237-9. [DOI] [PubMed] [Google Scholar]
  9. Jones B. N., Gurd R. S. Ultrapure cyanogen bromide-cleaved glucagon: isolation in high yield by ion-exchange chromatography. Arch Biochem Biophys. 1981 Jan;206(1):227–231. doi: 10.1016/0003-9861(81)90085-0. [DOI] [PubMed] [Google Scholar]
  10. Jones B. N., Stern A. S., Lewis R. V., Kimura S., Stein S., Udenfriend S., Shively J. E. Structure of two adrenal polypeptides containing multiple enkephalin sequences. Arch Biochem Biophys. 1980 Oct 1;204(1):392–395. doi: 10.1016/0003-9861(80)90048-x. [DOI] [PubMed] [Google Scholar]
  11. Kangawa K., Matsuo H. alpha-Neo-endorphin : a "big" Leu-enkephalin with potent opiate activity from porcine hypothalami. Biochem Biophys Res Commun. 1979 Jan 15;86(1):153–160. doi: 10.1016/0006-291x(79)90394-2. [DOI] [PubMed] [Google Scholar]
  12. Kimura S., Lewis R. V., Stern A. S., Rossier J., Stein S., Udenfriend S. Probable precursors of [Leu]enkephalin and [Met]enkephalin in adrenal medulla: peptides of 3-5 kilodaltons. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1681–1685. doi: 10.1073/pnas.77.3.1681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lewis R. V., Fallon A., Stein S., Gibson K. D., Udenfriend S. Supports for reverse-phase high-performance liquid chromatography of large proteins. Anal Biochem. 1980 May 1;104(1):153–159. doi: 10.1016/0003-2697(80)90291-2. [DOI] [PubMed] [Google Scholar]
  14. Lewis R. V., Stein S., Gerber L. D., Rubinstein M., Udenfriend S. High molecular weight opioid-containing proteins in striatum. Proc Natl Acad Sci U S A. 1978 Aug;75(8):4021–4023. doi: 10.1073/pnas.75.8.4021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lewis R. V., Stern A. S., Kilpatrick D. L., Gerber L. D., Rossier J., Stein S., Udenfriend S. Marked increases in large enkephalin-containing polypeptides in the rat adrenal gland following denervation. J Neurosci. 1981 Jan;1(1):80–82. doi: 10.1523/JNEUROSCI.01-01-00080.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lewis R. V., Stern A. S., Kimura S., Rossier J., Brink L., Gerber L. D., Stein S., Udenfriend S. Opioid peptides and precursors in the adrenal medulla. Adv Biochem Psychopharmacol. 1980;22:167–179. [PubMed] [Google Scholar]
  17. Lewis R. V., Stern A. S., Kimura S., Rossier J., Stein S., Udenfriend S. An about 50,000-dalton protein in adrenal medulla: a common precursor of [Met]- and [Leu]enkephalin. Science. 1980 Jun 27;208(4451):1459–1461. doi: 10.1126/science.7384787. [DOI] [PubMed] [Google Scholar]
  18. Lewis R. V., Stern A. S., Rossier J., Stein S., Udenfriend S. Putative enkephalin precursors in bovine adrenal medulla. Biochem Biophys Res Commun. 1979 Aug 13;89(3):822–829. doi: 10.1016/0006-291x(79)91852-7. [DOI] [PubMed] [Google Scholar]
  19. Mizuno K., Minamino N., Kangawa K., Matsuo H. A new endogenous opioid peptide from bovine adrenal medulla: isolation and amino acid sequence of a dodecapeptide (BAM-12P). Biochem Biophys Res Commun. 1980 Aug 29;95(4):1482–1488. doi: 10.1016/s0006-291x(80)80064-7. [DOI] [PubMed] [Google Scholar]
  20. Rossier J., Trifaró J. M., Lewis R. V., Lee R. W., Stern A., Kimura S., Stein S., Udenfriend S. Studies with [35S]methionine indicate that the 22,000-dalton [Met]enkephalin-containing protein in chromaffin cells is a precursor of [Met]enkephalin. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6889–6891. doi: 10.1073/pnas.77.11.6889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stein S., Böhlen P., Stone J., Dairman W., Udenfriend S. Amino acid analysis with fluorescamine at the picomole level. Arch Biochem Biophys. 1973 Mar;155(1):202–212. doi: 10.1016/s0003-9861(73)80022-0. [DOI] [PubMed] [Google Scholar]
  22. Steiner D. F., Quinn P. S., Chan S. J., Marsh J., Tager H. S. Processing mechanisms in the biosynthesis of proteins. Ann N Y Acad Sci. 1980;343:1–16. doi: 10.1111/j.1749-6632.1980.tb47238.x. [DOI] [PubMed] [Google Scholar]
  23. Stern A. S., Lewis R. V., Kimura S., Rossier J., Gerber L. D., Brink L., Stein S., Udenfriend S. Isolation of the opioid heptapeptide Met-enkephalin [Arg6,Phe7] from bovine adrenal medullary granules and striatum. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6680–6683. doi: 10.1073/pnas.76.12.6680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stern A. S., Lewis R. V., Kimura S., Rossier J., Stein S., Udenfriend S. Opioid hexapeptides and heptapeptides in adrenal medulla and brain possible implications on the biosynthesis of enkephalins. Arch Biochem Biophys. 1980 Dec;205(2):606–613. doi: 10.1016/0003-9861(80)90144-7. [DOI] [PubMed] [Google Scholar]
  25. Tager H. S., Emdin S. O., Clark J. L., Steiner D. F. Studies on the conversion of proinsulin to insulin. II. Evidence for a chymotrypsin-like cleavage in the connecting peptide region of insulin precursors in the rat. J Biol Chem. 1973 May 25;248(10):3476–3482. [PubMed] [Google Scholar]
  26. Wittmann-Liebold B. Amino acid sequence studies on ten ribosomal proteins of Escherichia coli with an improved sequenator equipped with an automatic conversion device. Hoppe Seylers Z Physiol Chem. 1973 Oct-Nov;354(10-11):1415–1431. doi: 10.1515/bchm2.1973.354.2.1415. [DOI] [PubMed] [Google Scholar]
  27. Yang H. Y., Fratta W., Hong J. S., DiGiulio A. M., Costa E. Detection of two endorphin-like peptides in nucleus caudatus. Neuropharmacology. 1978 Jun;17(6):433–438. doi: 10.1016/0028-3908(78)90023-0. [DOI] [PubMed] [Google Scholar]
  28. Zimmerman C. L., Appella E., Pisano J. J. Advances in the analysis of amino acid phenylthiohydantoins by high performance liquid chromatography. Anal Biochem. 1976 Sep;75(1):77–85. doi: 10.1016/0003-2697(76)90057-9. [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