Synthetic peptides comprising the amino-terminal sequence of a parathyroid hormone-like protein from human malignancies. Binding to parathyroid hormone receptors and activation of adenylate cyclase in bone cells and kidney

• Published in: The Journal of biological chemistry 1988-09, Vol.263 (26), p.12866-12871
• Main Authors: Nissenson, R A, Diep, D, Strewler, G J
• Format: Article
• Language: English
• Subjects: adenylate cyclase; Adenylyl Cyclases - metabolism; Amino Acid Sequence; Animals; Bone and Bones - enzymology; Dogs; Enzyme Activation; Guanylyl Imidodiphosphate - pharmacology; Humans; kidney; Kidney - enzymology; Neoplasm Proteins - analysis; osteosarcoma; parathyroid hormone; Parathyroid Hormone - analysis; Parathyroid Hormone - pharmacology; Parathyroid Hormone-Related Protein; Peptide Fragments - pharmacology; peptides; Receptors, Cell Surface - metabolism; Receptors, Parathyroid Hormone; Teriparatide - analogs & derivatives
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)37641-5

A tumor-derived protein with a spectrum of biologic activities remarkably similar to that of parathyroid hormone (PTH) has recently been purified and its sequence deduced from cloned cDNA. This PTH-like protein (PLP) has substantial sequence homology with PTH only in the amino-terminal 1-13 region and shows little similarity to other regions of PTH thought to be important for binding to receptors. In the present study, we compared the actions of two synthetic PLP peptides, PLP-(1-34)amide and [Tyr36]PLP-(1-36)amide, with those of bovine parathyroid hormone (bPTH)-(1-34) on receptors and adenylate cyclase in bone cells and in renal membranes. Synthetic PLP peptides were potent activators of adenylate cyclase in canine renal membranes (EC50 = 3.0 nM) and in UMR-106 osteosarcoma cells (EC50 = 0.05 nM). Bovine PTH-(1-34) was 6-fold more potent than the PLP peptides in renal membranes, but was 2-fold less potent in UMR-106 cells. A competitive PTH receptor antagonist, [Tyr34]bPTH-(7-34)amide, rapidly and fully inhibited adenylate cyclase stimulation by the PLP peptides as well as bPTH-(1-34). Competitive binding experiments with 125I-labeled PLP peptides revealed the presence of high affinity PLP receptors in UMR-106 cells IC50 = 3-4 nM) and in renal membranes (IC50 = 0.3 nM). There was no evidence of heterogeneity of PLP receptors. Bovine PTH-(1-34) was equipotent with the PLP peptides in binding to PLP receptors. Likewise, PLP peptides and bPTH-(1-34) were equipotent in competing with 125I-bPTH-(1-34) for binding to PTH receptors in renal membranes. Photoaffinity cross-linking experiments revealed that PTH and PLP peptides both interact with a major 85-kDa and minor 55- and 130-kDa components of canine renal membranes. We conclude that PTH and PLP activate adenylate cyclase by binding to common receptors in bone and kidney. The results further imply that subtle differences exist between PTH and PLP peptides in their ability to induce receptor-adenylate cyclase coupling.