RGD Reference Report - Single-channel function of recombinant type 2 inositol 1,4, 5-trisphosphate receptor. - Rat Genome Database

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Single-channel function of recombinant type 2 inositol 1,4, 5-trisphosphate receptor.

Authors: Ramos-Franco, J  Bare, D  Caenepeel, S  Nani, A  Fill, M  Mignery, G 
Citation: Ramos-Franco J, etal., Biophys J. 2000 Sep;79(3):1388-99.
RGD ID: 7175277
Pubmed: (View Article at PubMed) PMID:10969001
DOI: Full-text: DOI:10.1016/S0006-3495(00)76391-0

A full-length rat type 2 inositol 1,4,5-trisphosphate (InsP(3)) receptor cDNA construct was generated and expressed in COS-1 cells. Targeting of the full-length recombinant type 2 receptor protein to the endoplasmic reticulum was confirmed by immunocytochemistry using isoform specific affinity-purified antibodies and InsP(3)R-green fluorescent protein chimeras. The receptor protein was solubilized and incorporated into proteoliposomes for functional characterization. Single-channel recordings from proteoliposomes fused into planar lipid bilayers revealed that the recombinant protein formed InsP(3)- and Ca(2+)-sensitive ion channels. The unitary conductance ( approximately 250 pS; 220/20 mM Cs(+) as charge carrier), gating, InsP(3), and Ca(2+) sensitivities were similar to those previously described for the native type 2 InsP(3)R channel. However, the maximum open probability of the recombinant channel was slightly lower than that of its native counterpart. These data show that our full-length rat type 2 InsP(3)R cDNA construct encodes a protein that forms an ion channel with functional attributes like those of the native type 2 InsP(3)R channel. The possibility of measuring the function of single recombinant type 2 InsP(3)R is a significant step toward the use of molecular tools to define the determinants of isoform-specific InsP(3)R function and regulation.

Annotation

Gene Ontology Annotations    

Cellular Component

Molecular Function

Objects Annotated

Genes (Rattus norvegicus)
Itpr2  (inositol 1,4,5-trisphosphate receptor, type 2)


Additional Information