Purification and characterization of the inositol 1,4,5- trisphosphate receptor protein from rat vas deferens.
Among rat peripheral tissues examined, Ins(1,4,5)P(3) receptor binding is highest in the vas deferens, with levels about 25% of those of the cerebellum. We have purified the InsP(3) receptor binding protein from rat vas deferens membranes 600-fold. The purified protein displays a single 260 kDa band on SDS/PAGE, and the native protein has an apparent molecular mass of 1000 kDa, the same as in cerebellum. The inositol phosphate specificity, pH-dependence and influence of various reagents are the same for purified vas deferens and cerebellar receptors. Whereas particulate InsP(3) binding in cerebellum is potently inhibited by Ca(2+), particulate and purified vas deferens receptor binding of InsP(3) is not influenced by Ca(2+). Vas deferens appears to lack calmedin activity, but the InsP(3) receptor is sensitive to Ca(2+) inhibition conferred by brain calmedin. The vas deferens may prove to be a valuable tissue for characterizing functional aspects of InsP(3) receptors.
Mourey
R J
RJ
Johns Hopkins University School of Medicine, Department of Neuroscience, Baltimore, MD 21205.
Verma
A
A
Supattapone
S
S
Snyder
S H
SH
D000818:Animals; D001667:Binding, Competitive; D015220:Calcium Channels; D002462:Cell Membrane / Q000378:metabolism; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D053496:Inositol 1,4,5-Trisphosphate Receptors; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D017830:Octoxynol; D009928:Organ Specificity; D011092:Polyethylene Glycols; D051381:Rats; D011919:Rats, Inbred Strains; D011956:Receptors, Cell Surface / Q000302:isolation & purification* / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear; D014649:Vas Deferens / Q000378:metabolism*
D015220:Calcium Channels; D003902:Detergents; D053496:Inositol 1,4,5-Trisphosphate Receptors; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear; D011092:Polyethylene Glycols; D015544:Inositol 1,4,5-Trisphosphate; D017830:Octoxynol