Scaffolding proteins in highly purified rat olfactory cilia membranes

María Verónica Saavedra, Karl Heinz Smalla, Ulrich Thomas, Soledad Sandoval, Karel Olavarria, Karen Castillo, María Graciela Delgado, Ricardo Delgado, Eckart D. Gundelfinger, Juan Bacigalupo, Ursula Wyneken

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

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.
Original languageAmerican English
Pages (from-to)1123-1126
Number of pages4
JournalNeuroReport
Volume19
Issue number11
DOIs
StatePublished - 16 Jul 2008

Keywords

  • Olfactory epithelium
  • Scaffolding proteins
  • Signal transduction
  • Subcellular fractionation

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