Characterization of parathyroid hormone receptors in canine renal cortical plasma membranes using a radioiodinated sulfur-free hormone analogue. Correlation of binding with adenylate cyclase activity.

Segre, Gino V.; Rosenblatt, Michael; Reiner, B.; Mahaffey, J E; Potts, JT

doi:10.1016/s0021-9258(18)50271-4

Cited by 163 publications

(20 citation statements)

References 29 publications

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“…(A) Canine Renal Membranes. Purified canine renal cortical membranes were prepared in a fashion identical with that employed in an adenylate cyclase assay for PTH (Rosenblatt et al, 1977) and a radioligand-receptor binding assay for PTH (Segre et al, 1979) and were used previously in studies identifying PTH receptors by photoaffinity labeling (Coltrera et al, 1981;Draper et al, 1982).…”

Section: Methodsmentioning

confidence: 99%

“…The analogue can inhibit completely PTH-stimulated increases in cAMP in canine renal membranes (Rosenblatt et al, 1977), cells from human giant cell tumors, and human fibroblasts (Goldring et al, 1979). The analogue also has been shown to bind to PTH receptors with an avidity comparable to PTH (Segre et al, 1979) and to inhibit labeling of PTH receptors by a photolabile radiolabeled PTH analogue (Coltrera et al, 1981).…”

Section: Methodsmentioning

confidence: 99%

“…[Nle8,Nle18,Tyr34] bPTH-(1 -34)-amide was used for radiolabeling and reacted with photolabile groups as previously described (Coltrera et al, 1981). This analogue, previously synthesized, purified, and analyzed for chemical homogeneity and biological activity (Rosenblatt & Potts, 1977), was used as the radioligand for a PTH renal radioreceptor assay (Segre et al, 1979). [Nle8,Nle18,Tyr34]bPTH-(l-34)-amide (8 Mg) was iodinated with 6 mCi of Na125I (New England Nuclear) by a modification (Coltrera et al, 1981) of the method of Hunter & Greenwood (1962).…”

Section: Methodsmentioning

confidence: 99%

“…Desensitization to PTH requires more than 1 Abbreviations: PTH, parathyroid hormone; cAMP, adenosine cyclic 3',5'-phosphate; EDTA, ethylenediaminetetraacetate; bPTH, bovine parathyroid hormone; Tris, tris(hydroxymethyl)aminomethane; PBS, phosphate-buffered saline; NaDodS04, sodium dodecyl sulfate. simple occupancy of hormone receptors, since preincubation with a PTH analogue [which has been demonstrated to bind to PTH receptors (Segre et al, 1979) but which fails to stimulate a cAMP response (Rosenblatt et al, 1977;Goldring et al, 1979)] does not produce desensitization to PTH (Goldring et al, 1981). This result suggests that postbinding events, as postulated for other hormonal systems (Caro & Amatruda, 1980; Marshall & Olefsky, 1980;Su et al, 1980; King & Cuatrecasas, 1981), mediate hormone-specific desensitization.…”

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confidence: 99%

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Photoaffinity labeling of parathyroid hormone receptors: comparison of receptors across species and target tissues and after desensitization to hormone

Goldring¹,

Tyler²,

Krane³

et al. 1984

Biochemistry

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Cells derived from human giant cell tumors of bone and fibroblasts derived from human neonatal foreskin respond to parathyroid hormone (PTH) by increasing the intracellular and extracellular levels of adenosine cyclic 3',5'-phosphate (cAMP). Using photoaffinity labeling methods, we examined these cells for the presence of a PTH receptor or a binding subunit of a receptor complex. A previously designed biologically active and photolabile radioligand analogue of PTH was reacted with these intact cells. After photolysis, the cells were extracted, and the proteins were denatured, reduced, and separated by electrophoresis on sodium dodecyl sulfate (Na-DodSO4)-polyacrylamide gels followed by autoradiography. A single membrane component, Mr 70 000, was labeled specifically in intact cells cultured from skeletal and dermal tissue. By mixing, in pairs, photolabeled proteins from (a) intact human cells derived from giant cell tumors of bone, (b) intact human fibroblasts, and (c) canine renal cortical membranes, the receptors (or their binding subunits) for PTH were compared directly and found to be identical in terms of molecular size (as determined by the migration position on NaDod-SO4-polyacrylamide gels) across species (dog and human) and target tissue (bone, skin, and kidney). Preincubation of cells cultured from giant cell tumors of bone with PTH resulted in loss of the PTH-induced cAMP response (desensitization). Preincubation with PTH was accompanied by a marked decrease in photoaffinity labeling of the PTH binding component and suggests that the loss of hormone response in cells preincubated with PTH was related to a decrease in the number or availability of PTH receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Photoaffinity labeling of parathyroid hormone receptors: comparison of receptors across species and target tissues and after desensitization to hormone

Goldring¹,

Tyler²,

Krane³

et al. 1984

Biochemistry

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“…I^rathyroid hormone (PTH)1 receptors on target cells in bone and kidney initiate the biological effects of at least two humoral factors. PTH itself binds to receptors in bone (Pliam et al, 1982;Silve et al, 1982;Rizzoli et al, 1983a;Rao & Murray, 1985) and kidney (Zull et al, 1977;Nissenson & Arnaud, 1979;Segre et al, 1979;Bellorin-Font & Martin, 1981;Teitelbaum et al, 1982;Kremer et al, 1982;Forte et al, 1982; Rizzoli et al, 1983b;Teitelbaum & Strewler, 1984; McKee & , thereby activating the adenylate cyclase-cyclic AMP dependent protein kinase cascade, leading to cellular responses. In addition, investigators recently have identified a tumor-derived factor immunochemically distinct from PTH but capable of activating renal and skeletal PTH receptors (Strewler et al, 1983;Stewart et al, 1983;Rodan et al, 1983;Nissenson et al, 1985a).…”

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confidence: 99%

Covalent labeling of a high-affinity, guanyl nucleotide sensitive parathyroid hormone receptor in canine renal cortex

Nissenson¹,

Karpf²,

Bambino³

et al. 1987

Biochemistry

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Putative parathyroid hormone (PTH) receptors in canine renal membranes were affinity labeled with 125I-bPTH(1-34) using the heterobifunctional cross-linking reagent N-hydroxysuccinimidyl 4-azidobenzoate. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of a major 85,000 molecular weight (Mr) PTH binding component, the labeling of which was inhibited by nanomolar concentrations of unlabeled PTH and by micromolar concentrations of 5'-guanylyl imidodiphosphate [Gpp-(NH)p]. Labeling was not influenced by the unrelated peptides insulin and arginine vasopressin. Minor PTH binding components of Mr 55,000 and 130,000 were also seen, and labeling of these was likewise sensitive to unlabeled PTH and to Gpp(NH)p. Omission of protease inhibitors during the isolation of plasma membranes resulted in the loss of the Mr 85,000 PTH binding species and the appearance of an Mr 70,000 form. Several minor PTH binding components also were observed. Equilibrium binding studies showed that such membranes had an affinity for PTH indistinguishable from that in membranes isolated with protease inhibitors and displaying a major Mr 85,000 PTH binding species. We conclude that the major form of the adenylate cyclase coupled PTH receptor in canine renal membranes is an Mr 85,000 protein. An endogenous enzyme, probably a lysosomal cathepsin, can cleave this form to produce an Mr 70,000 receptor that retains full functional activity with respect to high-affinity, guanyl nucleotide sensitive PTH binding. The ability to covalently label the PTH receptor in high yield represents a major step toward the structural characterization of this important detector molecule.

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Two‐dimensional ¹H‐NMR study of the 1–34 fragment of human parathyroid hormone

Lee¹,

Russell²

1989

Biopolymers

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The complete assignment of resonances in the proton nmr spectrum of the 1-34 amino acid fragment of human parathyroid hormone [hPTH(1-34)], determined using a combination of one- and two-dimensional nmr techniques at 500 MHz, is described. In particular, homonuclear Hartmann-Hahn experiments, recorded in H2O and D2O, are used to resolve ambiguities in the connectivities between the highly overlapped resonances in the aliphatic region of the spectrum. One-dimensional multiple quantum filtering experiments are used to identify serine and phenylalanine spin systems. Analyses of the through-bond and through-space connectivities in the alpha H-NH fingerprint regions of the correlated spectroscopy (COSY) and nuclear Overhauser effect spectroscopy (NOESY) spectra lead to the assignment of resonances to specific amino acid residues in the polypeptide. Examination of the observed NOE cross peaks indicates that hPTH(1-34) has no detectable secondary structural elements in aqueous solution.

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Characterization of parathyroid hormone receptors in canine renal cortical plasma membranes using a radioiodinated sulfur-free hormone analogue. Correlation of binding with adenylate cyclase activity.

Cited by 163 publications

References 29 publications

Photoaffinity labeling of parathyroid hormone receptors: comparison of receptors across species and target tissues and after desensitization to hormone

Photoaffinity labeling of parathyroid hormone receptors: comparison of receptors across species and target tissues and after desensitization to hormone

Covalent labeling of a high-affinity, guanyl nucleotide sensitive parathyroid hormone receptor in canine renal cortex

Two‐dimensional ¹H‐NMR study of the 1–34 fragment of human parathyroid hormone

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Characterization of parathyroid hormone receptors in canine renal cortical plasma membranes using a radioiodinated sulfur-free hormone analogue. Correlation of binding with adenylate cyclase activity.

Cited by 163 publications

References 29 publications

Photoaffinity labeling of parathyroid hormone receptors: comparison of receptors across species and target tissues and after desensitization to hormone

Photoaffinity labeling of parathyroid hormone receptors: comparison of receptors across species and target tissues and after desensitization to hormone

Covalent labeling of a high-affinity, guanyl nucleotide sensitive parathyroid hormone receptor in canine renal cortex

Two‐dimensional 1H‐NMR study of the 1–34 fragment of human parathyroid hormone

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Two‐dimensional ¹H‐NMR study of the 1–34 fragment of human parathyroid hormone