title: Novel inactivating mutations of the calcium-sensing receptor: the calcimimetic NPS R-568 improves signal transduction of mutant receptors.
abstract: OBJECTIVE
  Inactivating mutations in the calcium-sensing receptor (CaSR) gene cause neonatal severe hyperparathyroidism and familial hypocalciuric hypercalcemia (FHH). The aims of the present study were the functional characterization of novel mutations of the CaSR found in FHH patients, the comparison of in vitro receptor function with clinical parameters, and the effect of the allosteric calcimimetic NPS R-568 on the signaling of mutant receptors.
  
  
  METHODS
  Wild-type and mutant CaSRs (W530G, C568Y, W718X, M734R, L849P, Q926R, and D1005N) were expressed in human embryonic kidney 293 cells. Receptor signaling was studied by measuring intracellular free calcium in response to different concentrations of extracellular calcium ([Ca(2+)](o)).
  
  
  RESULTS
  Four CaSR mutations (C568Y, W718X, M734R, and L849P) demonstrated a complete lack of a [Ca(2+)](o)-induced cytosolic Ca(2+) response up to 30 mm [Ca(2+)](o), whereas the CaSR mutants W530G, Q926R, and D1005N retained some sensitivity to [Ca(2+)](o). There was no significant relation between the in vitro calcium sensitivity, serum calcium, and intact PTH levels in the patients. Patients with C-terminal CaSR mutations had a calcium to creatine ratio above the established diagnostic threshold of 0.01 for FHH. The calcimimetic NPS R-568 enhanced the responsiveness to [Ca(2+)](o) in CaSR mutants of the extracellular domain (W530G and C568Y) as well as the intracellular C-terminal domain (Q926R and D1005N).
  
  
  CONCLUSIONS
  Therefore, calcimimetics might offer medical treatment for symptomatic FHH patients, and more important, for patients with neonatal severe hyperparathyroidism that harbor calcimimetic-sensitive CaSR mutants.
authors.affiliation: Division of Neuroendocrinology, Department of Neurosurgery, Friedrich-Alexander University Erlangen-Nuremberg, Schwabachanlage 6, 91054 Erlangen, Germany.
authors.forename: Ramona
authors.initials: R
authors.lastname: Rus
authors.forename: Christine
authors.initials: C
authors.lastname: Haag
authors.forename: Christiane
authors.initials: C
authors.lastname: Bumke-Vogt
authors.forename: Volker
authors.initials: V
authors.lastname: Bähr
authors.forename: Bernhard
authors.initials: B
authors.lastname: Mayr
authors.forename: Matthias
authors.initials: M
authors.lastname: Möhlig
authors.forename: Egbert
authors.initials: E
authors.lastname: Schulze
authors.forename: Karin
authors.initials: K
authors.lastname: Frank-Raue
authors.forename: Friedhelm
authors.initials: F
authors.lastname: Raue
authors.forename: Christof
authors.initials: C
authors.lastname: Schöfl
mesh_terms: D000814:Aniline Compounds / Q000494:pharmacology*; D015153:Blotting, Western; D002118:Calcium / Q000819:agonists / Q000097:blood; D002460:Cell Line; D006801:Humans; D006934:Hypercalcemia / Q000235:genetics / Q000503:physiopathology; D007011:Hypoparathyroidism / Q000235:genetics / Q000503:physiopathology; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010627:Phenethylamines; D011437:Propylamines; D044169:Receptors, Calcium-Sensing / Q000235:genetics* / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects* / Q000235:genetics*
chemical_list: D000814:Aniline Compounds; C095550:CASR protein, human; C107873:N-(2-chlorophenylpropyl)-1-(3-methoxyphenyl)ethylamine; D010627:Phenethylamines; D011437:Propylamines; D044169:Receptors, Calcium-Sensing; D002118:Calcium