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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
. 1982 Mar;79(6):2023–2025. doi: 10.1073/pnas.79.6.2023

Molecular forms of the putative enkephalin precursor BAM-12P in bovine adrenal, pituitary, and hypothalamus.

A Baird, N Ling, P Böhlen, R Benoit, R Klepper, R Guillemin
PMCID: PMC346114  PMID: 6952250

Abstract

A highly specific radioimmunoassay for one of the putative adrenomedullary [Met]enkephalin precursors, BAM-12P (Tyr-Gly-Gly-Phe-Met-Arg-Arg-Val-Gly-Arg-Pro-Glu-OH), has been developed. The BAM-12P antibodies recognize the COOH-terminal fragment of the peptide from Arg7 to Glu12 and do not crossreact with [Met5]- or [Leu5]enkephalin or any of their COOH-terminal lysine or arginine extended analogs. Specificity for the COOH-terminal Glu-OH is suggested by the 100% crossreactivity with BAM-12P5-12 and 0.3% crossreactivity with BAM-12P5-12 amide. Using these antibodies, we have measured three forms of BAM-12P-like immunoreactivity in extracts of bovine adrenal medulla, of which the major form (greater than 90%) corresponds to BAM-12P by molecular weight. Extracts of bovine adrenal cortex contain 1% the amount of a BAM-12P-like material (Mr approximately 1400; 20 ng per gland), possibly due to crosscontamination with adrenomedullary tissue. The major form of BAM-12P-like material in extracts of bovine neurointermediate pituitaries is of higher molecular weight than authentic BAM-12P (Mr approximately 4000); the remaining material (10%) corresponds to BAM-12P by molecular weight. There is no detectable BAM-12P-like immunoreactivity in crude or purified extracts of bovine anterior pituitaries. Extracts of bovine hypothalamic tissues contain small amounts of BAM-12P immunoreactivity (approximately 2 ng per fragment) which can be detected as one molecular form corresponding to a 1400-dalton molecule. The results indicate that the enkephalin precursor found in the adrenal medulla also may be present in the pituitary and hypothalamus. Furthermore, the processing of this molecule appears to be tissue-specific.

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Selected References

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

  1. Benoit R., Böhlen P., Ling N., Briskin A., Esch F., Brazeau P., Ying S. Y., Guillemin R. Presence of somatostatin-28-(1-12) in hypothalamus and pancreas. Proc Natl Acad Sci U S A. 1982 Feb;79(3):917–921. doi: 10.1073/pnas.79.3.917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  3. Kilpatrick D. L., Taniguchi T., Jones B. N., Stern A. S., Shively J. E., Hullihan J., Kimura S., Stein S., Udenfriend S. A highly potent 3200-dalton adrenal opioid peptide that contains both a [Met]- and [Leu]enkephalin sequence. Proc Natl Acad Sci U S A. 1981 May;78(5):3265–3268. doi: 10.1073/pnas.78.5.3265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Lewis R. V., Stern A. S., Kimura S., Stein S., Udenfriend S. Enkephalin biosynthetic pathway: proteins of 8000 and 14,000 daltons in bovine adrenal medulla. Proc Natl Acad Sci U S A. 1980 Aug;77(8):5018–5020. doi: 10.1073/pnas.77.8.5018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ling N. Solid phase synthesis of porcine alpha-endorphin and gamma-endorphin, two hypothalamic-pituitary peptides with opiate activity. Biochem Biophys Res Commun. 1977 Jan 10;74(1):248–255. doi: 10.1016/0006-291x(77)91401-2. [DOI] [PubMed] [Google Scholar]
  8. Lundberg J. M., Hamberger B., Schultzberg M., Hökfelt T., Granberg P. O., Efendić S., Terenius L., Goldstein M., Luft R. Enkephalin- and somatostatin-like immunoreactivities in human adrenal medulla and pheochromocytoma. Proc Natl Acad Sci U S A. 1979 Aug;76(8):4079–4083. doi: 10.1073/pnas.76.8.4079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Mizuno K., Minamino N., Kangawa K., Matsuo H. A new family of endogenous "big" Met-enkephalins from bovine adrenal medulla: purification and structure of docosa- (BAM-22P) and eicosapeptide (BAM-20P) with very potent opiate activity. Biochem Biophys Res Commun. 1980 Dec 31;97(4):1283–1290. doi: 10.1016/s0006-291x(80)80005-2. [DOI] [PubMed] [Google Scholar]
  11. Stern A. S., Jones B. N., Shively J. E., Stein S., Undenfriend S. Two adrenal opioid polypeptides: proposed intermediates in the processing of proenkephalin. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1962–1966. doi: 10.1073/pnas.78.3.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]

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