Mechanism-based labeling defines the free energy change for formation of the covalent glycosyl-enzyme intermediate in a xyloglucan endo-transglycosylase.
Identifieur interne : 003867 ( Main/Exploration ); précédent : 003866; suivant : 003868Mechanism-based labeling defines the free energy change for formation of the covalent glycosyl-enzyme intermediate in a xyloglucan endo-transglycosylase.
Auteurs : Kathleen Piens [Suède] ; Régis Fauré ; Gustav Sundqvist ; Martin J. Baumann ; Marc Saura-Valls ; Tuula T. Teeri ; Sylvain Cottaz ; Antoni Planas ; Hugues Driguez ; Harry BrumerSource :
- The Journal of biological chemistry [ 0021-9258 ] ; 2008.
Descripteurs français
- KwdFr :
- Coloration et marquage (MeSH), Glycosyltransferase (biosynthèse), Glycosyltransferase (composition chimique), Glycosyltransferase (métabolisme), Hydrolyse (MeSH), Oligosaccharides (composition chimique), Oligosaccharides (métabolisme), Populus (composition chimique), Populus (enzymologie), Spectrométrie de masse (MeSH), Thermodynamique (MeSH).
- MESH :
- biosynthèse : Glycosyltransferase.
- composition chimique : Glycosyltransferase, Oligosaccharides, Populus.
- enzymologie : Populus.
- métabolisme : Glycosyltransferase, Oligosaccharides.
- Coloration et marquage, Hydrolyse, Spectrométrie de masse, Thermodynamique.
English descriptors
- KwdEn :
- Glycosyltransferases (biosynthesis), Glycosyltransferases (chemistry), Glycosyltransferases (metabolism), Hydrolysis (MeSH), Mass Spectrometry (MeSH), Oligosaccharides (chemistry), Oligosaccharides (metabolism), Populus (chemistry), Populus (enzymology), Staining and Labeling (MeSH), Thermodynamics (MeSH).
- MESH :
- chemical , biosynthesis : Glycosyltransferases.
- chemical , chemistry : Glycosyltransferases, Oligosaccharides.
- chemical , metabolism : Glycosyltransferases, Oligosaccharides.
- chemistry : Populus.
- enzymology : Populus.
- Hydrolysis, Mass Spectrometry, Staining and Labeling, Thermodynamics.
Abstract
Xyloglucan endo-transglycosylases (XETs) are key enzymes involved in the restructuring of plant cell walls during morphogenesis. As members of glycoside hydrolase family 16 (GH16), XETs are predicted to employ the canonical retaining mechanism of glycosyl transfer involving a covalent glycosyl-enzyme intermediate. Here, we report the accumulation and direct observation of such intermediates of PttXET16-34 from hybrid aspen by electrospray mass spectrometry in combination with synthetic "blocked" substrates, which function as glycosyl donors but are incapable of acting as glycosyl acceptors. Thus, GalGXXXGGG and GalGXXXGXXXG react with the wild-type enzyme to yield relatively stable, kinetically competent, covalent GalG-enzyme and GalGXXXG-enzyme complexes, respectively (Gal=Galbeta(1-->4), G=Glcbeta(1-->4), and X=Xylalpha(1-->6)Glcbeta(1-->4)). Quantitation of ratios of protein and saccharide species at pseudo-equilibrium allowed us to estimate the free energy change (DeltaG(0)) for the formation of the covalent GalGXXXG-enzyme as 6.3-8.5 kJ/mol (1.5-2.0 kcal/mol). The data indicate that the free energy of the beta(1-->4) glucosidic bond in xyloglucans is preserved in the glycosyl-enzyme intermediate and harnessed for religation of the polysaccharide in vivo.
DOI: 10.1074/jbc.M803057200
PubMed: 18508770
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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Teeri</name></author><author><name sortKey="Cottaz, Sylvain" sort="Cottaz, Sylvain" uniqKey="Cottaz S" first="Sylvain" last="Cottaz">Sylvain Cottaz</name></author><author><name sortKey="Planas, Antoni" sort="Planas, Antoni" uniqKey="Planas A" first="Antoni" last="Planas">Antoni Planas</name></author><author><name sortKey="Driguez, Hugues" sort="Driguez, Hugues" uniqKey="Driguez H" first="Hugues" last="Driguez">Hugues Driguez</name></author><author><name sortKey="Brumer, Harry" sort="Brumer, Harry" uniqKey="Brumer H" first="Harry" last="Brumer">Harry Brumer</name></author></analytic><series><title level="j">The Journal of biological chemistry</title><idno type="ISSN">0021-9258</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Glycosyltransferases (biosynthesis)</term><term>Glycosyltransferases (chemistry)</term><term>Glycosyltransferases (metabolism)</term><term>Hydrolysis (MeSH)</term><term>Mass Spectrometry (MeSH)</term><term>Oligosaccharides (chemistry)</term><term>Oligosaccharides (metabolism)</term><term>Populus (chemistry)</term><term>Populus (enzymology)</term><term>Staining and Labeling (MeSH)</term><term>Thermodynamics (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Coloration et marquage (MeSH)</term><term>Glycosyltransferase (biosynthèse)</term><term>Glycosyltransferase (composition chimique)</term><term>Glycosyltransferase (métabolisme)</term><term>Hydrolyse (MeSH)</term><term>Oligosaccharides (composition chimique)</term><term>Oligosaccharides (métabolisme)</term><term>Populus (composition chimique)</term><term>Populus (enzymologie)</term><term>Spectrométrie de masse (MeSH)</term><term>Thermodynamique (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Glycosyltransferases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Glycosyltransferases</term><term>Oligosaccharides</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Glycosyltransferases</term><term>Oligosaccharides</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Glycosyltransferase</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Populus</term></keywords><keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Glycosyltransferase</term><term>Oligosaccharides</term><term>Populus</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Populus</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Populus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Glycosyltransferase</term><term>Oligosaccharides</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Hydrolysis</term><term>Mass Spectrometry</term><term>Staining and Labeling</term><term>Thermodynamics</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Coloration et marquage</term><term>Hydrolyse</term><term>Spectrométrie de masse</term><term>Thermodynamique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Xyloglucan endo-transglycosylases (XETs) are key enzymes involved in the restructuring of plant cell walls during morphogenesis. As members of glycoside hydrolase family 16 (GH16), XETs are predicted to employ the canonical retaining mechanism of glycosyl transfer involving a covalent glycosyl-enzyme intermediate. Here, we report the accumulation and direct observation of such intermediates of PttXET16-34 from hybrid aspen by electrospray mass spectrometry in combination with synthetic "blocked" substrates, which function as glycosyl donors but are incapable of acting as glycosyl acceptors. Thus, GalGXXXGGG and GalGXXXGXXXG react with the wild-type enzyme to yield relatively stable, kinetically competent, covalent GalG-enzyme and GalGXXXG-enzyme complexes, respectively (Gal=Galbeta(1-->4), G=Glcbeta(1-->4), and X=Xylalpha(1-->6)Glcbeta(1-->4)). Quantitation of ratios of protein and saccharide species at pseudo-equilibrium allowed us to estimate the free energy change (DeltaG(0)) for the formation of the covalent GalGXXXG-enzyme as 6.3-8.5 kJ/mol (1.5-2.0 kcal/mol). The data indicate that the free energy of the beta(1-->4) glucosidic bond in xyloglucans is preserved in the glycosyl-enzyme intermediate and harnessed for religation of the polysaccharide in vivo.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">18508770</PMID><DateCompleted><Year>2008</Year><Month>09</Month><Day>17</Day></DateCompleted><DateRevised><Year>2008</Year><Month>08</Month><Day>04</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Print">0021-9258</ISSN><JournalIssue CitedMedium="Print"><Volume>283</Volume><Issue>32</Issue><PubDate><Year>2008</Year><Month>Aug</Month><Day>08</Day></PubDate></JournalIssue><Title>The Journal of biological chemistry</Title><ISOAbbreviation>J Biol Chem</ISOAbbreviation></Journal><ArticleTitle>Mechanism-based labeling defines the free energy change for formation of the covalent glycosyl-enzyme intermediate in a xyloglucan endo-transglycosylase.</ArticleTitle><Pagination><MedlinePgn>21864-72</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1074/jbc.M803057200</ELocationID><Abstract><AbstractText>Xyloglucan endo-transglycosylases (XETs) are key enzymes involved in the restructuring of plant cell walls during morphogenesis. As members of glycoside hydrolase family 16 (GH16), XETs are predicted to employ the canonical retaining mechanism of glycosyl transfer involving a covalent glycosyl-enzyme intermediate. Here, we report the accumulation and direct observation of such intermediates of PttXET16-34 from hybrid aspen by electrospray mass spectrometry in combination with synthetic "blocked" substrates, which function as glycosyl donors but are incapable of acting as glycosyl acceptors. Thus, GalGXXXGGG and GalGXXXGXXXG react with the wild-type enzyme to yield relatively stable, kinetically competent, covalent GalG-enzyme and GalGXXXG-enzyme complexes, respectively (Gal=Galbeta(1-->4), G=Glcbeta(1-->4), and X=Xylalpha(1-->6)Glcbeta(1-->4)). Quantitation of ratios of protein and saccharide species at pseudo-equilibrium allowed us to estimate the free energy change (DeltaG(0)) for the formation of the covalent GalGXXXG-enzyme as 6.3-8.5 kJ/mol (1.5-2.0 kcal/mol). The data indicate that the free energy of the beta(1-->4) glucosidic bond in xyloglucans is preserved in the glycosyl-enzyme intermediate and harnessed for religation of the polysaccharide in vivo.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Piens</LastName><ForeName>Kathleen</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>School of Biotechnology, Royal Institute of Technology, AlbaNova University Centre, SE-106 91 Stockholm, Sweden.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Fauré</LastName><ForeName>Régis</ForeName><Initials>R</Initials></Author><Author ValidYN="Y"><LastName>Sundqvist</LastName><ForeName>Gustav</ForeName><Initials>G</Initials></Author><Author ValidYN="Y"><LastName>Baumann</LastName><ForeName>Martin J</ForeName><Initials>MJ</Initials></Author><Author ValidYN="Y"><LastName>Saura-Valls</LastName><ForeName>Marc</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Teeri</LastName><ForeName>Tuula T</ForeName><Initials>TT</Initials></Author><Author ValidYN="Y"><LastName>Cottaz</LastName><ForeName>Sylvain</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Planas</LastName><ForeName>Antoni</ForeName><Initials>A</Initials></Author><Author ValidYN="Y"><LastName>Driguez</LastName><ForeName>Hugues</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Brumer</LastName><ForeName>Harry</ForeName><Initials>H</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>2008</Year><Month>05</Month><Day>28</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>J Biol Chem</MedlineTA><NlmUniqueID>2985121R</NlmUniqueID><ISSNLinking>0021-9258</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D009844">Oligosaccharides</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 2.4.-</RegistryNumber><NameOfSubstance UI="D016695">Glycosyltransferases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 2.4.1.-</RegistryNumber><NameOfSubstance UI="C073693">xyloglucan endotransglycosylase</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D016695" MajorTopicYN="N">Glycosyltransferases</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006868" MajorTopicYN="N">Hydrolysis</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013058" MajorTopicYN="N">Mass Spectrometry</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009844" MajorTopicYN="N">Oligosaccharides</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D032107" MajorTopicYN="N">Populus</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013194" MajorTopicYN="N">Staining and Labeling</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013816" MajorTopicYN="Y">Thermodynamics</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2008</Year><Month>5</Month><Day>30</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2008</Year><Month>9</Month><Day>18</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2008</Year><Month>5</Month><Day>30</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">18508770</ArticleId><ArticleId IdType="pii">M803057200</ArticleId><ArticleId IdType="doi">10.1074/jbc.M803057200</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Suède</li></country></list><tree><noCountry><name sortKey="Baumann, Martin J" sort="Baumann, Martin J" uniqKey="Baumann M" first="Martin J" last="Baumann">Martin J. Baumann</name><name sortKey="Brumer, Harry" sort="Brumer, Harry" uniqKey="Brumer H" first="Harry" last="Brumer">Harry Brumer</name><name sortKey="Cottaz, Sylvain" sort="Cottaz, Sylvain" uniqKey="Cottaz S" first="Sylvain" last="Cottaz">Sylvain Cottaz</name><name sortKey="Driguez, Hugues" sort="Driguez, Hugues" uniqKey="Driguez H" first="Hugues" last="Driguez">Hugues Driguez</name><name sortKey="Faure, Regis" sort="Faure, Regis" uniqKey="Faure R" first="Régis" last="Fauré">Régis Fauré</name><name sortKey="Planas, Antoni" sort="Planas, Antoni" uniqKey="Planas A" first="Antoni" last="Planas">Antoni Planas</name><name sortKey="Saura Valls, Marc" sort="Saura Valls, Marc" uniqKey="Saura Valls M" first="Marc" last="Saura-Valls">Marc Saura-Valls</name><name sortKey="Sundqvist, Gustav" sort="Sundqvist, Gustav" uniqKey="Sundqvist G" first="Gustav" last="Sundqvist">Gustav Sundqvist</name><name sortKey="Teeri, Tuula T" sort="Teeri, Tuula T" uniqKey="Teeri T" first="Tuula T" last="Teeri">Tuula T. Teeri</name></noCountry><country name="Suède"><noRegion><name sortKey="Piens, Kathleen" sort="Piens, Kathleen" uniqKey="Piens K" first="Kathleen" last="Piens">Kathleen Piens</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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