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. 1999 Jul 15;341(Pt 2):271–276.

Identification of pituitary adenylate cyclase-activating polypeptide1-38-binding factor in human plasma, as ceruloplasmin.

J W Tams 1, A H Johnsen 1, J Fahrenkrug 1
PMCID: PMC1220356  PMID: 10393082

Abstract

125I-Pituitary adenylate cyclase-activating polypeptide (PACAP) 1-38 is able to bind a factor in human plasma, which can be displaced by unlabelled PACAP 1-38 and PACAP 28-38 but not by the other biologically active form, PACAP 1-27. Likewise, 125I-PACAP 28-38 binds this plasma factor, whereas 125I-PACAP 1-27 does not. Apparent Kd values were measured to be 12.0+/-1.3 and 3.4+/-1.5 nM for PACAP 1-38 and PACAP 28-38, respectively, using a competition assay with 125I-PACAP 28-38. Purification of the PACAP 1-38-binding factor from human blood was made by ethanol precipitation of serum followed by Ni2+-chelating and anion-exchange chromatography. A 120-kDa band on SDS/PAGE, as well as some proteolytic products, was blotted on to PVDF membrane and their N-terminal amino acid sequences determined. In combination with a mass-spectrometric fingerprinting of a tryptic digest of the 120-kDa band, this PACAP 1-38-binding factor was identified as ceruloplasmin. Purified commercial ceruloplasmin shows identical mobility on SDS/PAGE to the PACAP 1-38-binding factor and the same binding characteristics to PACAP 1-38, 1-27 and 28-38, using the same amount of ceruloplasmin as was expected to be found in the human plasma. Furthermore, the ability of plasma to bind 125I-PACAP 1-38 or 28-38 disappeared when ceruloplasmin was immunoprecipitated from plasma with rabbit anti-human ceruloplasmin Ig.

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Selected References

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  1. Aldred A. R., Grimes A., Schreiber G., Mercer J. F. Rat ceruloplasmin. Molecular cloning and gene expression in liver, choroid plexus, yolk sac, placenta, and testis. J Biol Chem. 1987 Feb 25;262(6):2875–2878. [PubMed] [Google Scholar]
  2. Arimura A., Shioda S. Pituitary adenylate cyclase activating polypeptide (PACAP) and its receptors: neuroendocrine and endocrine interaction. Front Neuroendocrinol. 1995 Jan;16(1):53–88. doi: 10.1006/frne.1995.1003. [DOI] [PubMed] [Google Scholar]
  3. Boutillier A. L., Monnier D., Koch B., Loeffler J. P. Pituitary adenyl cyclase-activating peptide: a hypophysiotropic factor that stimulates proopiomelanocortin gene transcription, and proopiomelanocortin-derived peptide secretion in corticotropic cells. Neuroendocrinology. 1994 Nov;60(5):493–502. doi: 10.1159/000126786. [DOI] [PubMed] [Google Scholar]
  4. Chiodera P., Volpi R., Capretti L., Caffarri G., Magotti M. G., Coiro V. Effects of intravenously infused pituitary adenylate cyclase-activating polypeptide on adenohypophyseal Hormone secretion in normal men. Neuroendocrinology. 1996 Sep;64(3):242–246. doi: 10.1159/000127124. [DOI] [PubMed] [Google Scholar]
  5. Fahrenkrug J., Hannibal J. Pituitary adenylate cyclase activating polypeptide immunoreactivity in capsaicin-sensitive nerve fibres supplying the rat urinary tract. Neuroscience. 1998 Apr;83(4):1261–1272. doi: 10.1016/s0306-4522(97)00474-0. [DOI] [PubMed] [Google Scholar]
  6. Fahrenkrug J., Hannibal J. Pituitary adenylate cyclase activating polypeptide innervation of the rat female reproductive tract and the associated paracervical ganglia: effect of capsaicin. Neuroscience. 1996 Aug;73(4):1049–1060. doi: 10.1016/0306-4522(96)00082-6. [DOI] [PubMed] [Google Scholar]
  7. Fleming R. E., Gitlin J. D. Primary structure of rat ceruloplasmin and analysis of tissue-specific gene expression during development. J Biol Chem. 1990 May 5;265(13):7701–7707. [PubMed] [Google Scholar]
  8. Frödin M., Hannibal J., Wulff B. S., Gammeltoft S., Fahrenkrug J. Neuronal localization of pituitary adenylate cyclase-activating polypeptide 38 in the adrenal medulla and growth-inhibitory effect on chromaffin cells. Neuroscience. 1995 Mar;65(2):599–608. doi: 10.1016/0306-4522(94)00522-7. [DOI] [PubMed] [Google Scholar]
  9. Gitlin J. D., Gitlin J. I., Gitlin D. Localizing of C-reactive protein in synovium of patients with rheumatoid arthritis. Arthritis Rheum. 1977 Nov-Dec;20(8):1491–1499. doi: 10.1002/art.1780200808. [DOI] [PubMed] [Google Scholar]
  10. Goth M. I., Lyons C. E., Canny B. J., Thorner M. O. Pituitary adenylate cyclase activating polypeptide, growth hormone (GH)-releasing peptide and GH-releasing hormone stimulate GH release through distinct pituitary receptors. Endocrinology. 1992 Feb;130(2):939–944. doi: 10.1210/endo.130.2.1346381. [DOI] [PubMed] [Google Scholar]
  11. Gräs S., Hannibal J., Georg B., Fahrenkrug J. Transient periovulatory expression of pituitary adenylate cyclase activating peptide in rat ovarian cells. Endocrinology. 1996 Nov;137(11):4779–4785. doi: 10.1210/endo.137.11.8895347. [DOI] [PubMed] [Google Scholar]
  12. Gutteridge J. M., Stocks J. Caeruloplasmin: physiological and pathological perspectives. Crit Rev Clin Lab Sci. 1981;14(4):257–329. doi: 10.3109/10408368109105866. [DOI] [PubMed] [Google Scholar]
  13. Hannibal J., Ekblad E., Mulder H., Sundler F., Fahrenkrug J. Pituitary adenylate cyclase activating polypeptide (PACAP) in the gastrointestinal tract of the rat: distribution and effects of capsaicin or denervation. Cell Tissue Res. 1998 Jan;291(1):65–79. doi: 10.1007/s004410050980. [DOI] [PubMed] [Google Scholar]
  14. Hannibal J., Fahrenkrug J. Expression of pituitary adenylate cyclase activating polypeptide (PACAP) gene by rat spermatogenic cells. Regul Pept. 1995 Jan 5;55(1):111–115. doi: 10.1016/0167-0115(94)00110-j. [DOI] [PubMed] [Google Scholar]
  15. Hannibal J., Mikkelsen J. D., Clausen H., Holst J. J., Wulff B. S., Fahrenkrug J. Gene expression of pituitary adenylate cyclase activating polypeptide (PACAP) in the rat hypothalamus. Regul Pept. 1995 Jan 26;55(2):133–148. doi: 10.1016/0167-0115(94)00099-j. [DOI] [PubMed] [Google Scholar]
  16. Hannibal J., Mikkelsen J. D., Fahrenkrug J., Larsen P. J. Pituitary adenylate cyclase-activating peptide gene expression in corticotropin-releasing factor-containing parvicellular neurons of the rat hypothalamic paraventricular nucleus is induced by colchicine, but not by adrenalectomy, acute osmotic, ether, or restraint stress. Endocrinology. 1995 Sep;136(9):4116–4124. doi: 10.1210/endo.136.9.7649120. [DOI] [PubMed] [Google Scholar]
  17. Hart G. R., Gowing H., Burrin J. M. Effects of a novel hypothalamic peptide, pituitary adenylate cyclase-activating polypeptide, on pituitary hormone release in rats. J Endocrinol. 1992 Jul;134(1):33–41. doi: 10.1677/joe.0.1340033. [DOI] [PubMed] [Google Scholar]
  18. Jaeger J. L., Shimizu N., Gitlin J. D. Tissue-specific ceruloplasmin gene expression in the mammary gland. Biochem J. 1991 Dec 15;280(Pt 3):671–677. doi: 10.1042/bj2800671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kononen J., Paavola M., Penttilä T. L., Parvinen M., Pelto-Huikko M. Stage-specific expression of pituitary adenylate cyclase-activating polypeptide (PACAP) mRNA in the rat seminiferous tubules. Endocrinology. 1994 Nov;135(5):2291–2294. doi: 10.1210/endo.135.5.7956953. [DOI] [PubMed] [Google Scholar]
  20. Martin J. L., Rose K., Hughes G. J., Magistretti P. J. [mono[125I]iodo-Tyr10,MetO17]-vasoactive intestinal polypeptide. Preparation, characterization, and use for radioimmunoassay and receptor binding. J Biol Chem. 1986 Apr 25;261(12):5320–5327. [PubMed] [Google Scholar]
  21. Mikkelsen J. D., Hannibal J., Fahrenkrug J., Larsen P. J., Olcese J., McArdle C. Pituitary adenylate cyclase activating peptide-38 (PACAP-38), PACAP-27, and PACAP related peptide (PRP) in the rat median eminence and pituitary. J Neuroendocrinol. 1995 Jan;7(1):47–55. doi: 10.1111/j.1365-2826.1995.tb00666.x. [DOI] [PubMed] [Google Scholar]
  22. Mikkelsen J. D., Hannibal J., Larsen P. J., Fahrenkrug J. Pituitary adenylate cyclase activating peptide (PACAP) mRNA in the rat neocortex. Neurosci Lett. 1994 Apr 25;171(1-2):121–124. doi: 10.1016/0304-3940(94)90620-3. [DOI] [PubMed] [Google Scholar]
  23. Miyata A., Arimura A., Dahl R. R., Minamino N., Uehara A., Jiang L., Culler M. D., Coy D. H. Isolation of a novel 38 residue-hypothalamic polypeptide which stimulates adenylate cyclase in pituitary cells. Biochem Biophys Res Commun. 1989 Oct 16;164(1):567–574. doi: 10.1016/0006-291x(89)91757-9. [DOI] [PubMed] [Google Scholar]
  24. Miyata A., Jiang L., Dahl R. D., Kitada C., Kubo K., Fujino M., Minamino N., Arimura A. Isolation of a neuropeptide corresponding to the N-terminal 27 residues of the pituitary adenylate cyclase activating polypeptide with 38 residues (PACAP38). Biochem Biophys Res Commun. 1990 Jul 31;170(2):643–648. doi: 10.1016/0006-291x(90)92140-u. [DOI] [PubMed] [Google Scholar]
  25. Nielsen H. S., Hannibal J., Fahrenkrug J. Expression of pituitary adenylate cyclase activating polypeptide (PACAP) in the postnatal and adult rat cerebellar cortex. Neuroreport. 1998 Aug 3;9(11):2639–2642. doi: 10.1097/00001756-199808030-00039. [DOI] [PubMed] [Google Scholar]
  26. Patel B. N., David S. A novel glycosylphosphatidylinositol-anchored form of ceruloplasmin is expressed by mammalian astrocytes. J Biol Chem. 1997 Aug 8;272(32):20185–20190. doi: 10.1074/jbc.272.32.20185. [DOI] [PubMed] [Google Scholar]
  27. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  28. Shioda S., Legradi G., Leung W. C., Nakajo S., Nakaya K., Arimura A. Localization of pituitary adenylate cyclase-activating polypeptide and its messenger ribonucleic acid in the rat testis by light and electron microscopic immunocytochemistry and in situ hybridization. Endocrinology. 1994 Sep;135(3):818–825. doi: 10.1210/endo.135.3.8070375. [DOI] [PubMed] [Google Scholar]
  29. Shvartsman A. L., Voronina O. V., Gaitskhoki V. S., Patkin E. L. Ekspressiia gena tseruloplazmina v organakh mlekopitaiushchikh po dannym gibridizatsionnogo analiza s komplementarnymi DNK-zondami. Mol Biol (Mosk) 1990 May-Jun;24(3):657–662. [PubMed] [Google Scholar]
  30. Steenstrup B. R., Alm P., Hannibal J., Jørgensen J. C., Palle C., Junge J., Christensen H. B., Ottesen B., Fahrenkrug J. Pituitary adenylate cyclase-activating polypeptide: occurrence and relaxant effect in female genital tract. Am J Physiol. 1995 Jul;269(1 Pt 1):E108–E117. doi: 10.1152/ajpendo.1995.269.1.E108. [DOI] [PubMed] [Google Scholar]
  31. Takahashi N., Ortel T. L., Putnam F. W. Single-chain structure of human ceruloplasmin: the complete amino acid sequence of the whole molecule. Proc Natl Acad Sci U S A. 1984 Jan;81(2):390–394. doi: 10.1073/pnas.81.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Thomas T., Schreiber G. The expression of genes coding for positive acute-phase proteins in the reproductive tract of the female rat. High levels of ceruloplasmin mRNA in the uterus. FEBS Lett. 1989 Jan 30;243(2):381–384. doi: 10.1016/0014-5793(89)80166-8. [DOI] [PubMed] [Google Scholar]
  33. Waschek J. A., Casillas R. A., Nguyen T. B., DiCicco-Bloom E. M., Carpenter E. M., Rodriguez W. I. Neural tube expression of pituitary adenylate cyclase-activating peptide (PACAP) and receptor: potential role in patterning and neurogenesis. Proc Natl Acad Sci U S A. 1998 Aug 4;95(16):9602–9607. doi: 10.1073/pnas.95.16.9602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yang F. M., Friedrichs W. E., Cupples R. L., Bonifacio M. J., Sanford J. A., Horton W. A., Bowman B. H. Human ceruloplasmin. Tissue-specific expression of transcripts produced by alternative splicing. J Biol Chem. 1990 Jun 25;265(18):10780–10785. [PubMed] [Google Scholar]
  35. Zhang Y. Z., Hannibal J., Zhao Q., Moller K., Danielsen N., Fahrenkrug J., Sundler F. Pituitary adenylate cyclase activating peptide expression in the rat dorsal root ganglia: up-regulation after peripheral nerve injury. Neuroscience. 1996 Oct;74(4):1099–1110. doi: 10.1016/0306-4522(96)00168-6. [DOI] [PubMed] [Google Scholar]

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