Human Ca2+ receptor extracellular domain. Analysis of function of lobe I loop deletion mutants

Guadalupe Reyes-Cruz, Jianxin Hu, Paul K. Goldsmith, Peter J. Steinbach, Allen M. Spiegel

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The G protein-coupled Ca2+ receptor (CaR) possesses an ∼600-residue extracellular domain involved in ligand binding and receptor activation. Based on an alignment of the amino acid sequence of the CaR with that of bacterial periplasmic-binding proteins, the first ∼530 residues of the extracellular domain are believed to form a domain resembling a bilobed Venus's flytrap (VFT). Four insertions in the CaR sequence that do not align with those of bacterial periplasmic-binding proteins correspond to four loops within lobe I of the VFT. We constructed a series of deletion mutants of these four loops and tested their ability to form fully processed CaR as well as their ability to be activated by Ca2+. As many as 21 residues (365-385) of loop III could be deleted without impairing receptor expression or activation. Deletion of portions of either loops I (50-59) or IV (438-445) did not impair receptor expression but significantly reduced Ca2+ activation. Deletion of the entire loop II (117-137) abolished receptor expression and function, but the replacement of even a single residue within this deletion mutant led to expression of a monomeric form of the receptor showing increased Ca2+ sensitivity but reduced maximal activation. Our results reveal that certain residues within loops I and IV are dispensable in formation of the VFT domain but are critical for Ca2+ activation of the receptor. In contrast, the residues in loop II are critical for maintaining the inactive state of the CaR. We discuss these results in light of the recently defined crystal structure of the homologous domain of the type 1 metabotropic glutamate receptor.

Original languageEnglish (US)
Pages (from-to)32145-32151
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number34
DOIs
StatePublished - Aug 24 2001
Externally publishedYes

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Droseraceae
Periplasmic Binding Proteins
Bacterial Proteins
Chemical activation
G-Protein-Coupled Receptors
Amino Acid Sequence
Ligands
GTP-Binding Proteins
Crystal structure
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Human Ca2+ receptor extracellular domain. Analysis of function of lobe I loop deletion mutants. / Reyes-Cruz, Guadalupe; Hu, Jianxin; Goldsmith, Paul K.; Steinbach, Peter J.; Spiegel, Allen M.

In: Journal of Biological Chemistry, Vol. 276, No. 34, 24.08.2001, p. 32145-32151.

Research output: Contribution to journalArticle

Reyes-Cruz, Guadalupe ; Hu, Jianxin ; Goldsmith, Paul K. ; Steinbach, Peter J. ; Spiegel, Allen M. / Human Ca2+ receptor extracellular domain. Analysis of function of lobe I loop deletion mutants. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 34. pp. 32145-32151.
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