Biophysical properties of the extra-cellular domain of the calcium-sensing receptor

Zachary C. Ryan, Theodore A. Craig, Sergei Yu Venyaminov, James R. Thompson, Rajiv Kumar

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The Calcium-Sensing Receptor (CaSR) is a G-protein-coupled receptor that regulates calcium homeostasis by altering parathyroid hormone release, and which binds divalent and trivalent cations, amino acids, polyamines, and polycationic ligands. To obtain information about the structural properties of the CaSR, we expressed milligram quantities of a pure, homogeneous, and functional fragment of the human CaSR extracellular domain (residues 20-535). The expressed and purified protein is folded and binds both neomycin and calcium. It forms dimers in the absence of reducing agents such as β-mercaptoethanol. Thermal denaturation studies show it has enthalpy and entropy values of unfolding equal to ΔH = -178 ± 4 kJ/mol and ΔS = -535 ± 13 J/mol/K. The protein has significant secondary structure with α-helical, β-sheet, β-turns, and disordered content of 36.6 ± 6.7%, 13.3 ± 5.3%, 20.2 ± 3.3%, and 29.4 ± 4.0%, respectively. The described method for the expression and purification of CaSR should prove useful for further structural studies of this physiologically important protein.

Original languageEnglish (US)
Pages (from-to)339-344
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume349
Issue number1
DOIs
StatePublished - Oct 13 2006

Fingerprint

Calcium-Sensing Receptors
Calcium
Denaturation
Neomycin
Mercaptoethanol
Reducing Agents
Divalent Cations
Polyamines
Entropy
G-Protein-Coupled Receptors
Parathyroid Hormone
Dimers
Purification
Cations
Structural properties
Enthalpy
Proteins
Homeostasis
Hot Temperature
Ligands

Keywords

  • Calcium
  • Calcium-sensing receptor
  • G-protein-coupled receptor
  • Secondary structure
  • Stability

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Biophysical properties of the extra-cellular domain of the calcium-sensing receptor. / Ryan, Zachary C.; Craig, Theodore A.; Venyaminov, Sergei Yu; Thompson, James R.; Kumar, Rajiv.

In: Biochemical and Biophysical Research Communications, Vol. 349, No. 1, 13.10.2006, p. 339-344.

Research output: Contribution to journalArticle

Ryan, Zachary C. ; Craig, Theodore A. ; Venyaminov, Sergei Yu ; Thompson, James R. ; Kumar, Rajiv. / Biophysical properties of the extra-cellular domain of the calcium-sensing receptor. In: Biochemical and Biophysical Research Communications. 2006 ; Vol. 349, No. 1. pp. 339-344.
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