Abstract
We have demonstrated binding of synthetic bovine parathyroid hormone (1-34) [bPTH-(1-34)] to embryonic avian bone cells in monolayer culture. The binding sites have qualitative and quantitative characteristics of a physiologically important parathyroid hormone (PTH) receptor. At apparent steady state (60 min at 24 degrees C), 5-10% of electrolytically labeled, receptor-purified 125I-labeled bPTH-(1-34) bound specifically to the cells whereas nonspecific binding was less than 1% of the added labeled hormone. Scatchard analysis showed a single order of PTH binding sites (Kd = 0.6 nM) with approximately 10,000 sites per cell. In this bone cell system, PTH bound to its binding site and stimulated cAMP accumulation over the same concentration range. Bovine PTH-(1-84) bound to the cells with the same apparent affinity as bPTH-(1-34).
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Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Brewer H. B., Jr, Fairwell T., Ronan R., Sizemore G. W., Arnaud C. D. Human parathyroid hormone: amino-acid sequence of the amino-terminal residues 1-34. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3585–3588. doi: 10.1073/pnas.69.12.3585. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Di Bella F. P., Dousa T. P., Miller S. S., Arnaud C. D. Parathyroid hormone receptors of renal cortex: specific binding of biologically active, 125I-labeled hormone and relationship to adenylate cyclase activation. Proc Natl Acad Sci U S A. 1974 Mar;71(3):723–726. doi: 10.1073/pnas.71.3.723. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gilman A. G. A protein binding assay for adenosine 3':5'-cyclic monophosphate. Proc Natl Acad Sci U S A. 1970 Sep;67(1):305–312. doi: 10.1073/pnas.67.1.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kugai N., Dorantes L., Nissenson R., Arnaud C. D. Parathyroid hormone receptors and stimulation of renal cyclic 3', 5' AMP in vitro: physiological relevance? Adv Exp Med Biol. 1978;103:537–542. doi: 10.1007/978-1-4684-7758-0_54. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Martin K. J., Hruska K. A., Freitag J. J., Klahr S., Slatopolsky E. The peripheral metabolism of parathyroid hormone. N Engl J Med. 1979 Nov 15;301(20):1092–1098. doi: 10.1056/NEJM197911153012005. [DOI] [PubMed] [Google Scholar]
- Nijweide P. J., van der Plas A., Scherft J. P. Biochemical and histological studies on various bone cell preparations. Calcif Tissue Int. 1981;33(5):529–540. doi: 10.1007/BF02409485. [DOI] [PubMed] [Google Scholar]
- Nissenson R. A., Arnaud C. D. Properties of the parathyroid hormone receptor-adenylate cyclase system in chicken renal plasma membranes. J Biol Chem. 1979 Mar 10;254(5):1469–1475. [PubMed] [Google Scholar]
- PECK W. A., BIRGE S. J., Jr, FEDAK S. A. BONE CELLS: BIOCHEMICAL AND BIOLOGICAL STUDIES AFTER ENZYMATIC ISOLATION. Science. 1964 Dec 11;146(3650):1476–1477. doi: 10.1126/science.146.3650.1476. [DOI] [PubMed] [Google Scholar]
- Peck W. A., Klahr S. Cyclic nucleotides in bone and mineral metabolism. Adv Cyclic Nucleotide Res. 1979;11:89–130. [PubMed] [Google Scholar]
- Wong G. L., Cohn D. V. Target cells in bone for parathormone and calcitonin are different: enrichment for each cell type by sequential digestion of mouse calvaria and selective adhesion to polymeric surfaces. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3167–3171. doi: 10.1073/pnas.72.8.3167. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zull J. E., Malbon C. C., Chuang J. Binding of tritiated bovine parathyroid hormone to plasma membranes from bovine kidney cortex. J Biol Chem. 1977 Feb 10;252(3):1071–1078. [PubMed] [Google Scholar]