An optimized method to extract poplar leaf proteins for two-dimensional gel electrophoresis guided by analysis of polysaccharides and phenolic compounds.
Identifieur interne : 002800 ( Main/Exploration ); précédent : 002799; suivant : 002801An optimized method to extract poplar leaf proteins for two-dimensional gel electrophoresis guided by analysis of polysaccharides and phenolic compounds.
Auteurs : Agnieszka Szuba [Pologne] ; Anna Wojakowska ; Gabriela Lorenc-Pluci SkaSource :
- Electrophoresis [ 1522-2683 ] ; 2013.
Descripteurs français
- KwdFr :
- MESH :
- analyse : Phénols, Polyosides, Protéines végétales.
- composition chimique : Feuilles de plante, Populus.
- isolement et purification : Protéines végétales.
- méthodes : Protéomique, Électrophorèse bidimensionnelle sur gel.
English descriptors
- KwdEn :
- MESH :
- chemical , analysis : Phenols, Plant Proteins, Polysaccharides.
- chemistry : Plant Leaves, Populus.
- chemical , isolation & purification : Plant Proteins.
- methods : Electrophoresis, Gel, Two-Dimensional, Proteomics.
Abstract
Commonly used methods for protein extraction from plant leaves, such as extraction with phenol or a combination of TCA and acetone, were ineffective for four tested cultivars of poplar. Moreover, multiple protocols for 2DE of the extracted proteins gave different results when protein profiles of relatively closely related plants were compared. Given that polycyclic compounds strongly hinder 2DE, we analyzed the impact of polyphenols and polysaccharides present in the plant tissues used for protein extraction, on the quality of 2DE protein profiles. Analysis of content of polyphenols and polysaccharides in leaves of poplar cultivars showed that even small differences in concentrations of analyzed metabolites accompany large differences between poplar cultivars when considering the susceptibility of samples to protein extraction for 2DE. High-quality 2DE results were correlated with decreased amounts of polyphenols. Additional analysis using MS/MS suggested that only levels of total phenolics affected the results of 2DE. Soluble total nonstructural carbohydrates also had a negative effect, but the level of starch was not important. Finally, we present an optimized method (OPTI) for extraction of proteins from poplar leaves, which enables reliable comparative analysis of four different poplar cultivars i.e. 'Eridano', 'Villafranca', 'NE-42' and 'Luisa Avanzo', which have not yet been used for the proteomic studies.
DOI: 10.1002/elps.201300223
PubMed: 26034795
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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gel</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Commonly used methods for protein extraction from plant leaves, such as extraction with phenol or a combination of TCA and acetone, were ineffective for four tested cultivars of poplar. Moreover, multiple protocols for 2DE of the extracted proteins gave different results when protein profiles of relatively closely related plants were compared. Given that polycyclic compounds strongly hinder 2DE, we analyzed the impact of polyphenols and polysaccharides present in the plant tissues used for protein extraction, on the quality of 2DE protein profiles. Analysis of content of polyphenols and polysaccharides in leaves of poplar cultivars showed that even small differences in concentrations of analyzed metabolites accompany large differences between poplar cultivars when considering the susceptibility of samples to protein extraction for 2DE. High-quality 2DE results were correlated with decreased amounts of polyphenols. Additional analysis using MS/MS suggested that only levels of total phenolics affected the results of 2DE. Soluble total nonstructural carbohydrates also had a negative effect, but the level of starch was not important. Finally, we present an optimized method (OPTI) for extraction of proteins from poplar leaves, which enables reliable comparative analysis of four different poplar cultivars i.e. 'Eridano', 'Villafranca', 'NE-42' and 'Luisa Avanzo', which have not yet been used for the proteomic studies.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">26034795</PMID><DateCompleted><Year>2015</Year><Month>10</Month><Day>09</Day></DateCompleted><DateRevised><Year>2018</Year><Month>10</Month><Day>23</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1522-2683</ISSN><JournalIssue CitedMedium="Internet"><Volume>34</Volume><Issue>22-23</Issue><PubDate><Year>2013</Year><Month>Dec</Month></PubDate></JournalIssue><Title>Electrophoresis</Title><ISOAbbreviation>Electrophoresis</ISOAbbreviation></Journal><ArticleTitle>An optimized method to extract poplar leaf proteins for two-dimensional gel electrophoresis guided by analysis of polysaccharides and phenolic compounds.</ArticleTitle><Pagination><MedlinePgn>3234-43</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1002/elps.201300223</ELocationID><Abstract><AbstractText>Commonly used methods for protein extraction from plant leaves, such as extraction with phenol or a combination of TCA and acetone, were ineffective for four tested cultivars of poplar. Moreover, multiple protocols for 2DE of the extracted proteins gave different results when protein profiles of relatively closely related plants were compared. Given that polycyclic compounds strongly hinder 2DE, we analyzed the impact of polyphenols and polysaccharides present in the plant tissues used for protein extraction, on the quality of 2DE protein profiles. Analysis of content of polyphenols and polysaccharides in leaves of poplar cultivars showed that even small differences in concentrations of analyzed metabolites accompany large differences between poplar cultivars when considering the susceptibility of samples to protein extraction for 2DE. High-quality 2DE results were correlated with decreased amounts of polyphenols. Additional analysis using MS/MS suggested that only levels of total phenolics affected the results of 2DE. Soluble total nonstructural carbohydrates also had a negative effect, but the level of starch was not important. Finally, we present an optimized method (OPTI) for extraction of proteins from poplar leaves, which enables reliable comparative analysis of four different poplar cultivars i.e. 'Eridano', 'Villafranca', 'NE-42' and 'Luisa Avanzo', which have not yet been used for the proteomic studies.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Szuba</LastName><ForeName>Agnieszka</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Laboratory of Proteomics, Institute of Dendrology, Polish Academy of Sciences, K órnik, Poland.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wojakowska</LastName><ForeName>Anna</ForeName><Initials>A</Initials></Author><Author ValidYN="Y"><LastName>Lorenc-Plucińska</LastName><ForeName>Gabriela</ForeName><Initials>G</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2013</Year><Month>10</Month><Day>01</Day></ArticleDate></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>Electrophoresis</MedlineTA><NlmUniqueID>8204476</NlmUniqueID><ISSNLinking>0173-0835</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010636">Phenols</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010940">Plant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011134">Polysaccharides</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D015180" MajorTopicYN="N">Electrophoresis, Gel, Two-Dimensional</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010636" MajorTopicYN="N">Phenols</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="Y">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018515" MajorTopicYN="N">Plant Leaves</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010940" MajorTopicYN="N">Plant Proteins</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011134" MajorTopicYN="N">Polysaccharides</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="Y">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D032107" MajorTopicYN="N">Populus</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D040901" MajorTopicYN="N">Proteomics</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2015</Year><Month>6</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2013</Year><Month>12</Month><Day>1</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2015</Year><Month>10</Month><Day>10</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">26034795</ArticleId><ArticleId IdType="doi">10.1002/elps.201300223</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Pologne</li></country></list><tree><noCountry><name sortKey="Lorenc Pluci Ska, Gabriela" sort="Lorenc Pluci Ska, Gabriela" uniqKey="Lorenc Pluci Ska G" first="Gabriela" last="Lorenc-Pluci Ska">Gabriela Lorenc-Pluci Ska</name><name sortKey="Wojakowska, Anna" sort="Wojakowska, Anna" uniqKey="Wojakowska A" first="Anna" last="Wojakowska">Anna Wojakowska</name></noCountry><country name="Pologne"><noRegion><name sortKey="Szuba, Agnieszka" sort="Szuba, Agnieszka" uniqKey="Szuba A" first="Agnieszka" last="Szuba">Agnieszka Szuba</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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