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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Comparison of Protein-Extraction Methods for Gills of the Shore Crab, <italic>Carcinus maenas</italic> (L.), and Application to 2DE</title><author><name sortKey="Panchout, Francois" sort="Panchout, Francois" uniqKey="Panchout F" first="François" last="Panchout">François Panchout</name><affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff></affiliation></author><author><name sortKey="Letendre, Julie" sort="Letendre, Julie" uniqKey="Letendre J" first="Julie" last="Letendre">Julie Letendre</name><affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff></affiliation></author><author><name sortKey="Bultelle, Florence" sort="Bultelle, Florence" uniqKey="Bultelle F" first="Florence" last="Bultelle">Florence Bultelle</name><affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff></affiliation></author><author><name sortKey="Denier, Xavier" sort="Denier, Xavier" uniqKey="Denier X" first="Xavier" last="Denier">Xavier Denier</name><affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff></affiliation></author><author><name sortKey="Rocher, Beatrice" sort="Rocher, Beatrice" uniqKey="Rocher B" first="Béatrice" last="Rocher">Béatrice Rocher</name><affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff></affiliation></author><author><name sortKey="Chan, Philippe" sort="Chan, Philippe" uniqKey="Chan P" first="Philippe" last="Chan">Philippe Chan</name><affiliation><nlm:aff wicri:cut=", and" id="aff3">Plate-forme de Protéomique Pissaro, Institute for Research and Innovation in Biomedicine</nlm:aff></affiliation></author><author><name sortKey="Vaudry, David" sort="Vaudry, David" uniqKey="Vaudry D" first="David" last="Vaudry">David Vaudry</name><affiliation><nlm:aff wicri:cut=", and" id="aff3">Plate-forme de Protéomique Pissaro, Institute for Research and Innovation in Biomedicine</nlm:aff></affiliation><affiliation><nlm:aff id="aff4">Neuronal and Neuroendocrine Differentiation and Communication, DC2N, Inserm U982, University of Rouen, 76821 Mont-Saint-Aignan, France</nlm:aff></affiliation></author><author><name sortKey="Durand, Fabrice" sort="Durand, Fabrice" uniqKey="Durand F" first="Fabrice" last="Durand">Fabrice Durand</name><affiliation><nlm:aff wicri:cut="; and" id="aff2">Faculty of Sciences and Technologies, University of Le Havre, 76058 Le Havre cedex, France</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">24294114</idno><idno type="pmc">3792702</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3792702</idno><idno type="RBID">PMC:3792702</idno><idno type="doi">10.7171/jbt.13-2404-002</idno><date when="2013">2013</date><idno type="wicri:Area/Pmc/Corpus">000186</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Comparison of Protein-Extraction Methods for Gills of the Shore Crab, <italic>Carcinus maenas</italic> (L.), and Application to 2DE</title><author><name sortKey="Panchout, Francois" sort="Panchout, Francois" uniqKey="Panchout F" first="François" last="Panchout">François Panchout</name><affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff></affiliation></author><author><name sortKey="Letendre, Julie" sort="Letendre, Julie" uniqKey="Letendre J" first="Julie" last="Letendre">Julie Letendre</name><affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff></affiliation></author><author><name sortKey="Bultelle, Florence" sort="Bultelle, Florence" uniqKey="Bultelle F" first="Florence" last="Bultelle">Florence Bultelle</name><affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff></affiliation></author><author><name sortKey="Denier, Xavier" sort="Denier, Xavier" uniqKey="Denier X" first="Xavier" last="Denier">Xavier Denier</name><affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff></affiliation></author><author><name sortKey="Rocher, Beatrice" sort="Rocher, Beatrice" uniqKey="Rocher B" first="Béatrice" last="Rocher">Béatrice Rocher</name><affiliation><nlm:aff wicri:cut=", and" id="aff1">Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222</nlm:aff></affiliation></author><author><name sortKey="Chan, Philippe" sort="Chan, Philippe" uniqKey="Chan P" first="Philippe" last="Chan">Philippe Chan</name><affiliation><nlm:aff wicri:cut=", and" id="aff3">Plate-forme de Protéomique Pissaro, Institute for Research and Innovation in Biomedicine</nlm:aff></affiliation></author><author><name sortKey="Vaudry, David" sort="Vaudry, David" uniqKey="Vaudry D" first="David" last="Vaudry">David Vaudry</name><affiliation><nlm:aff wicri:cut=", and" id="aff3">Plate-forme de Protéomique Pissaro, Institute for Research and Innovation in Biomedicine</nlm:aff></affiliation><affiliation><nlm:aff id="aff4">Neuronal and Neuroendocrine Differentiation and Communication, DC2N, Inserm U982, University of Rouen, 76821 Mont-Saint-Aignan, France</nlm:aff></affiliation></author><author><name sortKey="Durand, Fabrice" sort="Durand, Fabrice" uniqKey="Durand F" first="Fabrice" last="Durand">Fabrice Durand</name><affiliation><nlm:aff wicri:cut="; and" id="aff2">Faculty of Sciences and Technologies, University of Le Havre, 76058 Le Havre cedex, France</nlm:aff></affiliation></author></analytic><series><title level="j">Journal of Biomolecular Techniques : JBT</title><idno type="ISSN">1524-0215</idno><idno type="eISSN">1943-4731</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>As it is well-established that protein extraction constitutes a crucial step for two-dimensional electrophoresis (2DE), this work was done as a prerequisite to further the study of alterations in the proteome in gills of the shore crab <italic>Carcinus maenas</italic> under contrasted environmental conditions. Because of the presence of a chitin layer, shore crab gills have an unusual structure. Consequently, they are considered as a hard tissue and represent a challenge for optimal protein extraction. In this study, we compared three published extraction procedures for subsequent applications to 2DE: the first one uses homogenization process, the second one included an additional TCA-acetone precipitation step, and finally, the third one associated grinding in liquid nitrogen (N<sub>2</sub>) and TCA-acetone precipitation. Extracted proteins were then resolved using 1DE and 2DE. Although interesting patterns were obtained using 1DE with the three methods, only the one involving grinding in liquid N<sub>2</sub> and TCA-acetone precipitation led to proper resolution after 2DE, showing a good level of reproducibility at technical (85%) and biological (84%) levels. This last method is therefore proposed for analysis of gill proteomes in the shore crab.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Biomol Tech</journal-id><journal-id journal-id-type="iso-abbrev">J Biomol Tech</journal-id><journal-id journal-id-type="pmc">jbt</journal-id><journal-id journal-id-type="publisher-id">jbt</journal-id><journal-title-group><journal-title>Journal of Biomolecular Techniques : JBT</journal-title></journal-title-group><issn pub-type="ppub">1524-0215</issn><issn pub-type="epub">1943-4731</issn><publisher><publisher-name>Association of Biomolecular Resource Facilities</publisher-name><publisher-loc>Bethesda, MD</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">24294114</article-id><article-id pub-id-type="pmc">3792702</article-id><article-id pub-id-type="publisher-id">13-2404-002</article-id><article-id pub-id-type="doi">10.7171/jbt.13-2404-002</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Comparison of Protein-Extraction Methods for Gills of the Shore Crab, <italic>Carcinus maenas</italic> (L.), and Application to 2DE</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Panchout</surname><given-names>François</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref><xref ref-type="corresp" rid="cor1"></xref></contrib><contrib contrib-type="author"><name><surname>Letendre</surname><given-names>Julie</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Bultelle</surname><given-names>Florence</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Denier</surname><given-names>Xavier</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Rocher</surname><given-names>Béatrice</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Chan</surname><given-names>Philippe</given-names></name><xref ref-type="aff" rid="aff3"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Vaudry</surname><given-names>David</given-names></name><xref ref-type="aff" rid="aff3"><sup>2</sup></xref><xref ref-type="aff" rid="aff4"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Durand</surname><given-names>Fabrice</given-names></name><xref ref-type="aff" rid="aff2"><sup>4</sup></xref></contrib><aff id="aff1"><sup>1</sup>Laboratory of Ecotoxicology–Aquatic Environments, LEMA, EA 3222, and</aff><aff id="aff2"><sup>4</sup>Faculty of Sciences and Technologies, University of Le Havre, 76058 Le Havre cedex, France; and</aff><aff id="aff3"><sup>2</sup>Plate-forme de Protéomique Pissaro, Institute for Research and Innovation in Biomedicine, and</aff><aff id="aff4"><sup>3</sup>Neuronal and Neuroendocrine Differentiation and Communication, DC2N, Inserm U982, University of Rouen, 76821 Mont-Saint-Aignan, France</aff></contrib-group><author-notes><corresp id="cor1">A<sc>ddress correspondence to</sc>: François Panchout, <addr-line>Laboratory of Ecotoxicology–Aquatic Environments (LEMA, EA 3222), University of Le Havre, B.P. 540, 76058 Le Havre Cedex, France</addr-line> (Phone: <phone>+33(0)2 32 85 99 09</phone>; Fax: <fax>+33(0)2 32 74 45 05</fax>; E-mail: <email>francois.panchout@univ-lehavre.fr</email>).</corresp></author-notes><pub-date pub-type="ppub"><month>12</month><year>2013</year></pub-date><pub-date pub-type="pmc-release"><month>12</month><year>2013</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on the
<volume>24</volume><issue>4</issue><fpage>218</fpage><lpage>223</lpage><permissions><copyright-statement>© 2013 Association of Biomolecular Resource Facilities</copyright-statement><copyright-year>2013</copyright-year></permissions><self-uri xlink:title="pdf" xlink:type="simple" xlink:href="jbt00413000218.pdf"></self-uri><abstract><p>As it is well-established that protein extraction constitutes a crucial step for two-dimensional electrophoresis (2DE), this work was done as a prerequisite to further the study of alterations in the proteome in gills of the shore crab <italic>Carcinus maenas</italic> under contrasted environmental conditions. Because of the presence of a chitin layer, shore crab gills have an unusual structure. Consequently, they are considered as a hard tissue and represent a challenge for optimal protein extraction. In this study, we compared three published extraction procedures for subsequent applications to 2DE: the first one uses homogenization process, the second one included an additional TCA-acetone precipitation step, and finally, the third one associated grinding in liquid nitrogen (N<sub>2</sub>) and TCA-acetone precipitation. Extracted proteins were then resolved using 1DE and 2DE. Although interesting patterns were obtained using 1DE with the three methods, only the one involving grinding in liquid N<sub>2</sub> and TCA-acetone precipitation led to proper resolution after 2DE, showing a good level of reproducibility at technical (85%) and biological (84%) levels. This last method is therefore proposed for analysis of gill proteomes in the shore crab.</p></abstract><kwd-group><kwd>2D gel electrophoresis</kwd><kwd>hard tissue</kwd><kwd>chitin</kwd><kwd>crustacean</kwd></kwd-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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