Activation of Skeletal Adenyl Cyclase by Parathyroid Hormone<i>in Vitro</i><sup>1</sup>

Chase, Lewis R.; Fedak, Susan A.; Aurbach, G. D.

doi:10.1210/endo-84-4-761

Cited by 233 publications

(65 citation statements)

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“…Characterization of binding sites in the above systems indicate that they function as hormone receptors coupled to adenylate cyclase, consistent with the well-established response to PTH, i.e. production of adenosinc 3',5'-monophosphate (CAMP) [8,91. Both preparations of radiolabeled ligand [I, 21 have Icd to the development of photoaffinity probes which have identified a single membrane component, M , 70000, which may represent either the PTH receptor or a binding subunit of the receptor [lo-121.…”

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confidence: 71%

Identification of a 150‐kDa membrane component which is modulated by parathyroid hormone

Weinshank

Luben

1985

European Journal of Biochemistry

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Monoclonal antibodies have been produced against primary bone cells obtained from the collagenase digestion of mouse cranial bone. Antibodies were selected on the basis of their immunoglobulin class and those which were identified as IgC were further screened for their ability to inhibit CAMP accumulation in response to sub-maximal doses of the 1 -34 amino-terminal peptide of bovine parathyroid hormone, bPTH(1-34). Nine hybridoma clones were subsequently characterized as inhibitory with respect to parathyroid hormone (PTH) responses in intact mouse cranial bone and which also identified a variety of membrane components from detergent extracts of surface-labeled primary bone cells. Five of these antibodies immunoprecipitated a membrane component with M , of 80000 that appeared to be a major component of the extract susceptible to surface-labeling with lZ51. All nine monoclonal antibodies were shown to bind to a suspcndcd-cell preparation of primary bone cells with 2 -3 orders of magnitude greater binding than that of control antibodies. Using this assay, one clone, designated 3G12 IgG, was observed to exhibit desensitization effects at the binding level with a time course and dose dependency for PTH pre-incubation that was similar to the establishment of the refractory state in other systems. In addition, the desensitization effect oCcurred at 37°C but not at 4°C. This antibody was shown to bind saturably to both intact mouse cranial bone and primary bone cells with an apparent affinity constant (Ka) in the range of lo9 M. Inhibition of bone CAMP accumulation in response to 2.5 nM bPTH(1-34) was directly correlated to the binding of 3G12 IgC to intact mouse calvariae. A niaximuiii inhibition of approximately 85% was obscrved. 3G12 IgC iinmunoprecipitated a single membrane component, M , 150000, from NP-40 detergent extracts of "sI-labeled primary mouse bone cells. The molecular mass of this component was also 150000 daltons when run on polyacrylamide gel slabs under non-reducing conditions. Control and PTH-pre-treated bone cells were surfacelabeled, detergent-solubilized and immunoprecipitated with 3G12 IgG in order to investigate further the desensitization effect at the molecular level. Incubation of bone cells with 1 pg/ml bPTH(1 -34) for 45 min at 37 C caused an increased susceptibility to surface-labeling with IZ5I that was approximately three-fold higher in specific activity than that of control cells. Under conditions of quantitative iminunoprccipitation, 3GI 2 IgG precipitated less of the 150-kDa membrane component from the PTH pre-treated bone cell extract, indicating a decreased availability of this membrane component under conditions of desensitization. We propose that the 150-kDa protein from mouse bone cells may be the PTH receptor, a component of the receptor or a membrane protein which is directly involved in the PTH responsiveness of these cells.Parathyroid hormone (PTH) is a peptide hormone which initiates a variety of biological activities by binding to specific cell-surface receptors prcscnt on...

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confidence: 71%

Identification of a 150‐kDa membrane component which is modulated by parathyroid hormone

Weinshank

Luben

1985

European Journal of Biochemistry

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“…Removal of the thyroid glands from two patients with pseudohypoparathyroidism caused only a transient rise in serum calcium in one patient (36) and no detectable change in the other (39). Further, studies with rats show that thyrocalcitonin does not inhibit the activation of renal adenyl cyclase by parathyroid hormone, nor does it influence urinary excretion of 3',5'-AMP (2,5,12 Lee (35) showing that extracts from the parathyroid glands of a patient with pseudohypoparathyroidism caused a hypercalcemic response in parathyroidectomized test animals makes this thesis untenable. In the current study, induction of hypercalcemia with consequent inhibition of secretion of immunologically reactive parathyroid hormone did not correct the defective renal response to exogenous hormone.…”

Section: Discussionmentioning

confidence: 99%

“…This hypothesis was based on the observation that patients with this disorder did not show the characteristic phosphaturic response to parathyroid extract. The recent findings that the mechanism of action of parathyroid hormone is mediated through activation of adenyl cyclase in kidney (2)(3)(4) and bone (5) suggested further investigation into the nature of the biochemical defect in pseudohypoparathyroidism. Parathyroid hormone affects specifically the membrane-bound enzyme adenyl cyclase in the renal cortex (3) and in bone (5) causing a marked increase in the concentration of adenosine 3',5'-monophosphate (3',5'-AMP) in these tissues (6,71).…”

Section: Introductionmentioning

confidence: 99%

Pseudohypoparathyroidism: defective excretion of 3′,5′-AMP in response to parathyroid hormone

Chase¹,

Melson²,

Aurbach³

1969

J. Clin. Invest.

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468

158

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A B S T R A C T Urinary excretion of cyclic adenosine 3',5'-monophosphate (3',5'-AMP) was tested in normal subjects and patients with pseudohypoparathyroidism, idiopathic hypoparathyroidism, surgical hypoparathyroidism, and pseudopseudohypoparathyroidism under basal conditions and after a 15 min infusion of purified parathyroid hormone. Basal excretion of the nucleotide was less than normal in the patients with hypocalcemic disorders and greater than normal in pseudopseudohypoparathyroidism. Parathyroid hormone caused a marked increase in excretion of 3',5'-AMP in all subjects except those with pseudohypoparathyroidism; nine patients with this disorder did not respond to the hormone and four showed a markedly deficient response. Radioimmunoassay showed that parathyroid hormone circulated in increased amounts in plasma from patients with pseudohypoparathyroidism and became undetectable when serum calcium was increased above 12 mg/100 ml. Suppression of parathyroid hormone secretion by induction of hypercalcemia did not alter the deficient response to exogenous hormone. The results indicate that: (a) parathyroid hormone circulates in abnormally high concentrations in pseudohypoparathyroidism and secretion of the hormone responds normally to physiological control by calcium; (b) testing urinary excretion of 3',5'-AMP in response to infusion of purified parathyroid hormone appears to-be an accurate and sensitive index for establishing the diagnosis of pseudohypoparathyroidism; and (c) the metabolic defect of the disorder can be accounted for by a lack of or defective form of parathyroid hormone-sensitive adenyl cyclase in bone and kidney.

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“…The physiologic action of PTH on bone [6], kidney cortex [4,5,14,15,19], and renal tubules [14] is mediated through activation of adenyl cyclase, the enzyme which catalyzes the conversion of adenosine triphosphate to cyclic AMP. Activation of adenyl cyclase results in increased cyclic AMP in renal parenchyma [16,18] and increased cyclic AMP excretion in urine [1,2,4,8].…”

Section: Introductionmentioning

confidence: 99%

The Effect of Parathyroid Hormone on Rabbit Renal Cortex Adenyl Cyclase during Development

Linarelli

Bobik

1973

Pediatr Res

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ExtractRenal cortex from fetal and neonatal rabbits (from 4-5 days preterm to 4 days of age) shows results comparable with maternal renal cortex during in vitro stimulation of the 2,200 X g pellet with parathyroid hormone (PTH) at 20 /ug/ml. Incubation with sodium fluoride (10 HIM), a nonspecific potent stimulator of adenyl cyclase, resulted in comparable increases in both the fetal and the maternal renal cortex. Glucagon, prostaglandin Ei, and vasopressin, all at 20 jug/ml, stimulate adenyl cyclase to a lesser extent than PTH. Homogenates of full term neonatal rabbits and maternal renal cortex show comparable phosphodiesterase enzyme levels with or without 0.04 M imidazole. These data show that in fetal and early neonatal renal cortex there is no developmental adenyl cyclase responsiveness to PTH. Similarly, phosphodiesterase is present equally in term and maternal rabbit renal cortex. Speculation

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Activation of Skeletal Adenyl Cyclase by Parathyroid Hormonein Vitro¹

Cited by 233 publications

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Identification of a 150‐kDa membrane component which is modulated by parathyroid hormone

Identification of a 150‐kDa membrane component which is modulated by parathyroid hormone

Pseudohypoparathyroidism: defective excretion of 3′,5′-AMP in response to parathyroid hormone

The Effect of Parathyroid Hormone on Rabbit Renal Cortex Adenyl Cyclase during Development

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Activation of Skeletal Adenyl Cyclase by Parathyroid Hormonein Vitro1

Cited by 233 publications

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Identification of a 150‐kDa membrane component which is modulated by parathyroid hormone

Identification of a 150‐kDa membrane component which is modulated by parathyroid hormone

Pseudohypoparathyroidism: defective excretion of 3′,5′-AMP in response to parathyroid hormone

The Effect of Parathyroid Hormone on Rabbit Renal Cortex Adenyl Cyclase during Development

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Activation of Skeletal Adenyl Cyclase by Parathyroid Hormonein Vitro¹