Proteomic Analysis of a Membrane Preparation from Rat Olfactory Sensory Cilia
The cilia of mammalian olfactory receptor neurons (ORNs) represent the sensory interface that is exposed to the air within the nasal cavity. The cilia are the site where odorants bind to specific receptors and initiate olfactory transduction that leads to excitation of the neuron. This process invol...
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| Veröffentlicht in: | Chemical senses 2008-02, Vol.33 (2), p.145-162 |
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| creator | Mayer, Ulrich Ungerer, Nicole Klimmeck, Daniel Warnken, Uwe Schnölzer, Martina Frings, Stephan Möhrlen, Frank |
| description | The cilia of mammalian olfactory receptor neurons (ORNs) represent the sensory interface that is exposed to the air within the nasal cavity. The cilia are the site where odorants bind to specific receptors and initiate olfactory transduction that leads to excitation of the neuron. This process involves a multitude of ciliary proteins that mediate chemoelectrical transduction, amplification, and adaptation of the primary sensory signal. Many of these proteins were initially identified by their enzymatic activities using a membrane protein preparation from olfactory cilia. This so-called “calcium-shock” preparation is a versatile tool for the exploration of protein expression, enzyme kinetics, regulatory mechanisms, and ciliary development. To support such studies, we present a first proteomic analysis of this membrane preparation. We subjected the cilia preparation to liquid chromatography-electrospray ionisation (LC-ESI-MS/MS) tandem mass spectrometry and identified 268 proteins, of which 49% are membrane proteins. A detailed analysis of their cellular and subcellular localization showed that the cilia preparation obtained by calcium shock not only is highly enriched in ORN proteins but also contains a significant amount of nonciliary material. Although our proteomic study does not identify the entire set of ciliary and nonciliary proteins, it provides the first estimate of the purity of the calcium-shock preparation and provides valuable biochemical information for further research. |
| doi_str_mv | 10.1093/chemse/bjm073 |
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The cilia are the site where odorants bind to specific receptors and initiate olfactory transduction that leads to excitation of the neuron. This process involves a multitude of ciliary proteins that mediate chemoelectrical transduction, amplification, and adaptation of the primary sensory signal. Many of these proteins were initially identified by their enzymatic activities using a membrane protein preparation from olfactory cilia. This so-called “calcium-shock” preparation is a versatile tool for the exploration of protein expression, enzyme kinetics, regulatory mechanisms, and ciliary development. To support such studies, we present a first proteomic analysis of this membrane preparation. We subjected the cilia preparation to liquid chromatography-electrospray ionisation (LC-ESI-MS/MS) tandem mass spectrometry and identified 268 proteins, of which 49% are membrane proteins. A detailed analysis of their cellular and subcellular localization showed that the cilia preparation obtained by calcium shock not only is highly enriched in ORN proteins but also contains a significant amount of nonciliary material. Although our proteomic study does not identify the entire set of ciliary and nonciliary proteins, it provides the first estimate of the purity of the calcium-shock preparation and provides valuable biochemical information for further research.</description><identifier>ISSN: 0379-864X</identifier><identifier>EISSN: 1464-3553</identifier><identifier>DOI: 10.1093/chemse/bjm073</identifier><identifier>PMID: 18032372</identifier><identifier>CODEN: CHSED8</identifier><language>eng</language><publisher>Oxford: Oxford University Press</publisher><subject>Animals ; Biological and medical sciences ; Chromatography, Liquid ; Cilia - chemistry ; Computational Biology ; Fundamental and applied biological sciences. Psychology ; Membrane Proteins - analysis ; Membrane Proteins - isolation & purification ; Olfaction. Taste ; Olfactory Mucosa - chemistry ; Olfactory Mucosa - cytology ; olfactory receptor neurons ; Olfactory Receptor Neurons - chemistry ; Olfactory Receptor Neurons - ultrastructure ; Perception ; Proteins - analysis ; proteomic analysis ; Proteomics ; Psychology. Psychoanalysis. Psychiatry ; Psychology. Psychophysiology ; Rats ; Rats, Wistar ; sensory cilia ; signal transduction ; Smell ; Spectrometry, Mass, Electrospray Ionization</subject><ispartof>Chemical senses, 2008-02, Vol.33 (2), p.145-162</ispartof><rights>The Author 2007. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org 2008</rights><rights>2008 INIST-CNRS</rights><rights>The Author 2007. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c489t-b11e4ccc9447ad2e05e95585ec4bcb2f36dfb6debcc5a03a498b13385002aa393</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,1583,27922,27923</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=20082851$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18032372$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Mayer, Ulrich</creatorcontrib><creatorcontrib>Ungerer, Nicole</creatorcontrib><creatorcontrib>Klimmeck, Daniel</creatorcontrib><creatorcontrib>Warnken, Uwe</creatorcontrib><creatorcontrib>Schnölzer, Martina</creatorcontrib><creatorcontrib>Frings, Stephan</creatorcontrib><creatorcontrib>Möhrlen, Frank</creatorcontrib><title>Proteomic Analysis of a Membrane Preparation from Rat Olfactory Sensory Cilia</title><title>Chemical senses</title><addtitle>Chem Senses</addtitle><description>The cilia of mammalian olfactory receptor neurons (ORNs) represent the sensory interface that is exposed to the air within the nasal cavity. The cilia are the site where odorants bind to specific receptors and initiate olfactory transduction that leads to excitation of the neuron. This process involves a multitude of ciliary proteins that mediate chemoelectrical transduction, amplification, and adaptation of the primary sensory signal. Many of these proteins were initially identified by their enzymatic activities using a membrane protein preparation from olfactory cilia. This so-called “calcium-shock” preparation is a versatile tool for the exploration of protein expression, enzyme kinetics, regulatory mechanisms, and ciliary development. To support such studies, we present a first proteomic analysis of this membrane preparation. We subjected the cilia preparation to liquid chromatography-electrospray ionisation (LC-ESI-MS/MS) tandem mass spectrometry and identified 268 proteins, of which 49% are membrane proteins. A detailed analysis of their cellular and subcellular localization showed that the cilia preparation obtained by calcium shock not only is highly enriched in ORN proteins but also contains a significant amount of nonciliary material. Although our proteomic study does not identify the entire set of ciliary and nonciliary proteins, it provides the first estimate of the purity of the calcium-shock preparation and provides valuable biochemical information for further research.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Chromatography, Liquid</subject><subject>Cilia - chemistry</subject><subject>Computational Biology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Membrane Proteins - analysis</subject><subject>Membrane Proteins - isolation & purification</subject><subject>Olfaction. Taste</subject><subject>Olfactory Mucosa - chemistry</subject><subject>Olfactory Mucosa - cytology</subject><subject>olfactory receptor neurons</subject><subject>Olfactory Receptor Neurons - chemistry</subject><subject>Olfactory Receptor Neurons - ultrastructure</subject><subject>Perception</subject><subject>Proteins - analysis</subject><subject>proteomic analysis</subject><subject>Proteomics</subject><subject>Psychology. Psychoanalysis. Psychiatry</subject><subject>Psychology. Psychophysiology</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>sensory cilia</subject><subject>signal transduction</subject><subject>Smell</subject><subject>Spectrometry, Mass, Electrospray Ionization</subject><issn>0379-864X</issn><issn>1464-3553</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0cFrFDEUBvAgit1Wj14lCIqXsUleMpMc27VacUtrVShewks2g7PObNZkBtz_3llmqOClp3f58b3H-wh5wdk7zgyc-p-hy-HUbTpWwSOy4LKUBSgFj8mCQWUKXcq7I3Kc84YxLkHop-SIawYCKrEgVzcp9iF2jadnW2z3uck01hTpVehcwm2gNynsMGHfxC2tU-zoLfb0uq3R9zHt6dewzYe5bNoGn5EnNbY5PJ_nCfn-4eLb8rJYXX_8tDxbFV5q0xeO8yC990bKCtciMBWMUloFL513ooZyXbtyHZz3ChmgNNpxAK0YE4hg4IS8mXJ3Kf4eQu5t12Qf2nY8OA7ZVgwYNwYehIIpADCHxFf_wU0c0viRbMcgIThIPaJiQj7FnFOo7S41Haa95cwe2rBTG3ZqY_Qv59DBdWH9T8_vH8HrGWD22Nbjw32T751gTAut-OjeTi4Ouwd3zjc2uQ9_7jGmX7asoFL28u6Hvf3yfik_n6-sgb9mOLC4</recordid><startdate>20080201</startdate><enddate>20080201</enddate><creator>Mayer, Ulrich</creator><creator>Ungerer, Nicole</creator><creator>Klimmeck, Daniel</creator><creator>Warnken, Uwe</creator><creator>Schnölzer, Martina</creator><creator>Frings, Stephan</creator><creator>Möhrlen, Frank</creator><general>Oxford University Press</general><general>Oxford Publishing Limited (England)</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QG</scope><scope>7QP</scope><scope>7QR</scope><scope>7SS</scope><scope>7TK</scope><scope>8FD</scope><scope>FR3</scope><scope>K9.</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20080201</creationdate><title>Proteomic Analysis of a Membrane Preparation from Rat Olfactory Sensory Cilia</title><author>Mayer, Ulrich ; Ungerer, Nicole ; Klimmeck, Daniel ; Warnken, Uwe ; Schnölzer, Martina ; Frings, Stephan ; Möhrlen, Frank</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c489t-b11e4ccc9447ad2e05e95585ec4bcb2f36dfb6debcc5a03a498b13385002aa393</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Chromatography, Liquid</topic><topic>Cilia - chemistry</topic><topic>Computational Biology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Membrane Proteins - analysis</topic><topic>Membrane Proteins - isolation & purification</topic><topic>Olfaction. Taste</topic><topic>Olfactory Mucosa - chemistry</topic><topic>Olfactory Mucosa - cytology</topic><topic>olfactory receptor neurons</topic><topic>Olfactory Receptor Neurons - chemistry</topic><topic>Olfactory Receptor Neurons - ultrastructure</topic><topic>Perception</topic><topic>Proteins - analysis</topic><topic>proteomic analysis</topic><topic>Proteomics</topic><topic>Psychology. Psychoanalysis. Psychiatry</topic><topic>Psychology. Psychophysiology</topic><topic>Rats</topic><topic>Rats, Wistar</topic><topic>sensory cilia</topic><topic>signal transduction</topic><topic>Smell</topic><topic>Spectrometry, Mass, Electrospray Ionization</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mayer, Ulrich</creatorcontrib><creatorcontrib>Ungerer, Nicole</creatorcontrib><creatorcontrib>Klimmeck, Daniel</creatorcontrib><creatorcontrib>Warnken, Uwe</creatorcontrib><creatorcontrib>Schnölzer, Martina</creatorcontrib><creatorcontrib>Frings, Stephan</creatorcontrib><creatorcontrib>Möhrlen, Frank</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Neurosciences Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Chemical senses</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mayer, Ulrich</au><au>Ungerer, Nicole</au><au>Klimmeck, Daniel</au><au>Warnken, Uwe</au><au>Schnölzer, Martina</au><au>Frings, Stephan</au><au>Möhrlen, Frank</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Proteomic Analysis of a Membrane Preparation from Rat Olfactory Sensory Cilia</atitle><jtitle>Chemical senses</jtitle><addtitle>Chem Senses</addtitle><date>2008-02-01</date><risdate>2008</risdate><volume>33</volume><issue>2</issue><spage>145</spage><epage>162</epage><pages>145-162</pages><issn>0379-864X</issn><eissn>1464-3553</eissn><coden>CHSED8</coden><abstract>The cilia of mammalian olfactory receptor neurons (ORNs) represent the sensory interface that is exposed to the air within the nasal cavity. 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| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current); EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | Animals Biological and medical sciences Chromatography, Liquid Cilia - chemistry Computational Biology Fundamental and applied biological sciences. Psychology Membrane Proteins - analysis Membrane Proteins - isolation & purification Olfaction. Taste Olfactory Mucosa - chemistry Olfactory Mucosa - cytology olfactory receptor neurons Olfactory Receptor Neurons - chemistry Olfactory Receptor Neurons - ultrastructure Perception Proteins - analysis proteomic analysis Proteomics Psychology. Psychoanalysis. Psychiatry Psychology. Psychophysiology Rats Rats, Wistar sensory cilia signal transduction Smell Spectrometry, Mass, Electrospray Ionization |
| title | Proteomic Analysis of a Membrane Preparation from Rat Olfactory Sensory Cilia |
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