title: The membrane proteome of sensory cilia to the depth of olfactory receptors.
abstract: In the nasal cavity, the nonmotile cilium of olfactory sensory neurons (OSNs) constitutes the chemosensory interface between the ambient environment and the brain. The unique sensory organelle facilitates odor detection for which it includes all necessary components of initial and downstream olfactory signal transduction. In addition to its function in olfaction, a more universal role in modulating different signaling pathways is implicated, for example, in neurogenesis, apoptosis, and neural regeneration. To further extend our knowledge about this multifunctional signaling organelle, it is of high importance to establish a most detailed proteome map of the ciliary membrane compartment down to the level of transmembrane receptors. We detached cilia from mouse olfactory epithelia via Ca(2+)/K(+) shock followed by the enrichment of ciliary membrane proteins at alkaline pH, and we identified a total of 4,403 proteins by gel-based and gel-free methods in conjunction with high resolution LC/MS. This study is the first to report the detection of 62 native olfactory receptor proteins and to provide evidence for their heterogeneous expression at the protein level. Quantitative data evaluation revealed four ciliary membrane-associated candidate proteins (the annexins ANXA1, ANXA2, ANXA5, and S100A5) with a suggested function in the regulation of olfactory signal transduction, and their presence in ciliary structures was confirmed by immunohistochemistry. Moreover, we corroborated the ciliary localization of the potassium-dependent Na(+)/Ca(2+) exchanger (NCKX) 4 and the plasma membrane Ca(2+)-ATPase 1 (PMCA1) involved in olfactory signal termination, and we detected for the first time NCKX2 in olfactory cilia. Through comparison with transcriptome data specific for mature, ciliated OSNs, we finally delineated the membrane ciliome of OSNs. The membrane proteome of olfactory cilia established here is the most complete today, thus allowing us to pave new avenues for the study of diverse molecular functions and signaling pathways in and out of olfactory cilia and thus to advance our understanding of the biology of sensory organelles in general.
authors.affiliation: From the ‡Medizinisches Proteom-Center, Ruhr-Universität Bochum, Universitätsstrasse 150, 44780 Bochum.
authors.forename: Katja
authors.initials: K
authors.lastname: Kuhlmann
authors.affiliation: From the ‡Medizinisches Proteom-Center, Ruhr-Universität Bochum, Universitätsstrasse 150, 44780 Bochum.
authors.forename: Astrid
authors.initials: A
authors.lastname: Tschapek
authors.affiliation: the ¶Faculty of Biology and BIOSS Centre for Biological Signalling Studies, University of Freiburg, Schänzlestrasse 1, 79104 Freiburg.
authors.forename: Heike
authors.initials: H
authors.lastname: Wiese
authors.affiliation: From the ‡Medizinisches Proteom-Center, Ruhr-Universität Bochum, Universitätsstrasse 150, 44780 Bochum.
authors.forename: Martin
authors.initials: M
authors.lastname: Eisenacher
authors.affiliation: From the ‡Medizinisches Proteom-Center, Ruhr-Universität Bochum, Universitätsstrasse 150, 44780 Bochum, the ‖Leibniz-Institut für Analytische Wissenschaften-ISAS-e.V., Otto-Hahn-Strasse 6b, 44227 Dortmund, and.
authors.forename: Helmut E
authors.initials: HE
authors.lastname: Meyer
authors.affiliation: the **Department of Cell Physiology, Ruhr-Universität Bochum, Universitätsstrasse 150, 44780 Bochum, Germany.
authors.forename: Hanns H
authors.initials: HH
authors.lastname: Hatt
authors.affiliation: the ¶Faculty of Biology and BIOSS Centre for Biological Signalling Studies, University of Freiburg, Schänzlestrasse 1, 79104 Freiburg.
authors.forename: Silke
authors.initials: S
authors.lastname: Oeljeklaus
authors.affiliation: the ¶Faculty of Biology and BIOSS Centre for Biological Signalling Studies, University of Freiburg, Schänzlestrasse 1, 79104 Freiburg, bettina.warscheid@biologie.uni-freiburg.de.
authors.forename: Bettina
authors.initials: B
authors.lastname: Warscheid
mesh_terms: D000818:Animals; D017305:Annexin A1 / Q000378:metabolism; D017306:Annexin A2 / Q000378:metabolism; D017304:Annexin A5 / Q000378:metabolism; D017920:Antiporters / Q000378:metabolism; D002923:Cilia; D020869:Gene Expression Profiling; D008297:Male; D051379:Mice; D009296:Nasal Cavity / Q000294:innervation*; D009812:Odorants; D018034:Olfactory Receptor Neurons / Q000378:metabolism*; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000378:metabolism; D020543:Proteome / Q000032:analysis; D018035:Receptors, Odorant / Q000032:analysis* / Q000096:biosynthesis; D009418:S100 Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D012903:Smell / Q000502:physiology*; D019831:Sodium-Calcium Exchanger / Q000378:metabolism
chemical_list: D017305:Annexin A1; D017306:Annexin A2; D017304:Annexin A5; D017920:Antiporters; D020543:Proteome; D018035:Receptors, Odorant; D009418:S100 Proteins; C569957:Slc24a4 protein, mouse; D019831:Sodium-Calcium Exchanger; C074530:potassium-dependent sodium-calcium exchanger; D053499:Plasma Membrane Calcium-Transporting ATPases; C506954:Atp2b1 protein, mouse
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