TY - JOUR
T1 - Scaffolding proteins in highly purified rat olfactory cilia membranes
AU - Saavedra, María Verónica
AU - Smalla, Karl Heinz
AU - Thomas, Ulrich
AU - Sandoval, Soledad
AU - Olavarria, Karel
AU - Castillo, Karen
AU - Delgado, María Graciela
AU - Delgado, Ricardo
AU - Gundelfinger, Eckart D.
AU - Bacigalupo, Juan
AU - Wyneken, Ursula
PY - 2008/7/16
Y1 - 2008/7/16
N2 - Odour-mediated signal transduction is a complex process that occurs in the cilia of olfactory sensory neurons. To gain insight in to the molecular organization of the odour transduction machinery, we developed a procedure to purify olfactory cilia membranes by differential centrifugation of rat olfactory epithelium extracts. We tested whether known scaffolding proteins that might participate in the assembly of the complex chemotransduction apparatus are present in the purified membrane fraction. Utilizing immunoblotting and immunohistochemistry, we show that the multidomain scaffolding proteins ProSAP/Shanks and calcium/calmodulin-dependent serine protein kinase CASK are present in the olfactory cilia. Ion channels involved in chemotransduction could be reconstituted into planar lipid bilayers for electrophysiological recordings. Our procedure should allow the identification of further chemotransduction-related proteins.
AB - Odour-mediated signal transduction is a complex process that occurs in the cilia of olfactory sensory neurons. To gain insight in to the molecular organization of the odour transduction machinery, we developed a procedure to purify olfactory cilia membranes by differential centrifugation of rat olfactory epithelium extracts. We tested whether known scaffolding proteins that might participate in the assembly of the complex chemotransduction apparatus are present in the purified membrane fraction. Utilizing immunoblotting and immunohistochemistry, we show that the multidomain scaffolding proteins ProSAP/Shanks and calcium/calmodulin-dependent serine protein kinase CASK are present in the olfactory cilia. Ion channels involved in chemotransduction could be reconstituted into planar lipid bilayers for electrophysiological recordings. Our procedure should allow the identification of further chemotransduction-related proteins.
KW - Olfactory epithelium
KW - Scaffolding proteins
KW - Signal transduction
KW - Subcellular fractionation
KW - Olfactory epithelium
KW - Scaffolding proteins
KW - Signal transduction
KW - Subcellular fractionation
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UR - https://www.scopus.com/inward/citedby.uri?partnerID=HzOxMe3b&scp=49949106254&origin=inward
U2 - 10.1097/WNR.0b013e3283086797
DO - 10.1097/WNR.0b013e3283086797
M3 - Article
SN - 0959-4965
VL - 19
SP - 1123
EP - 1126
JO - NeuroReport
JF - NeuroReport
IS - 11
ER -