GlutaredoxinV1, Main, Exploration, bibRecord, 000598

Structure-guided activity enhancement and catalytic mechanism of yeast grx8.

Identifieur interne : 000598 ( Main/Exploration ); précédent : 000597; suivant : 000599

Structure-guided activity enhancement and catalytic mechanism of yeast grx8.

Auteurs : Yajun Tang [République populaire de Chine] ; Jiahai Zhang ; Jiang Yu ; Ling Xu ; Jihui Wu ; Cong-Zhao Zhou ; Yunyu Shi

Source :

Biochemistry [ 1520-4995 ] ; 2014.

RBID : pubmed:24611845

Descripteurs français

KwdFr :
- Activation enzymatique (MeSH), Biocatalyse (MeSH), Cinétique (MeSH), Conformation des protéines (MeSH), Glutarédoxines (composition chimique), Glutarédoxines (isolement et purification), Glutarédoxines (métabolisme), Protéines recombinantes (composition chimique), Protéines recombinantes (isolement et purification), Protéines recombinantes (métabolisme), Résonance magnétique nucléaire biomoléculaire (MeSH), Saccharomyces cerevisiae (enzymologie).
MESH :
- composition chimique : Glutarédoxines, Protéines recombinantes.
- enzymologie : Saccharomyces cerevisiae.
- isolement et purification : Glutarédoxines, Protéines recombinantes.
- métabolisme : Glutarédoxines, Protéines recombinantes.
- Activation enzymatique, Biocatalyse, Cinétique, Conformation des protéines, Résonance magnétique nucléaire biomoléculaire.

English descriptors

KwdEn :
- Biocatalysis (MeSH), Enzyme Activation (MeSH), Glutaredoxins (chemistry), Glutaredoxins (isolation & purification), Glutaredoxins (metabolism), Kinetics (MeSH), Nuclear Magnetic Resonance, Biomolecular (MeSH), Protein Conformation (MeSH), Recombinant Proteins (chemistry), Recombinant Proteins (isolation & purification), Recombinant Proteins (metabolism), Saccharomyces cerevisiae (enzymology).
MESH :
- chemical , chemistry : Glutaredoxins, Recombinant Proteins.
- chemical , isolation & purification : Glutaredoxins, Recombinant Proteins.
- chemical , metabolism : Glutaredoxins, Recombinant Proteins.
- enzymology : Saccharomyces cerevisiae.
- Biocatalysis, Enzyme Activation, Kinetics, Nuclear Magnetic Resonance, Biomolecular, Protein Conformation.

Abstract

Glutaredoxins (Grxs) are wide-spread oxidoreductases that are found in all kingdoms of life. The yeast Saccharomyces cerevisiae encodes eight Grxs, among which, Grx8 shares a sequence identity of 30 and 23% with typical dithiol Grx1 and Grx2, respectively, but it exhibits a much lower GSH-dependent oxidoreductase activity. To elucidate its catalytic mechanism, we solved the solution structure of Grx8, which displays a typical Grx fold. Structural analysis indicated that Grx8 possesses a negatively charged CXXC motif (Cys(33)-Pro(34)-Asp(35)-Cys(36)) and a GSH-recognition site, which are distinct from Grx1 and Grx2. Subsequent structure-guided site mutations revealed that the D35Y single mutant and N80T/L81V double mutant possess increased activity of 10- and 11-fold, respectively; moreover, the D35Y/N80T/L81V triple mutant has increased activity of up to 44-fold, which is comparable to that of canonical Grx. Biochemical analyses suggested that the increase in catalytic efficiency resulted from a decreased pKa value of catalytic cysteine Cys33 and/or enhancement of the putative GSH-recognition site. Moreover, NMR chemical shift perturbation analyses combined with GSH analogue inhibition assays enabled us to elucidate that wild-type Grx8 and all mutants adopt a ping-pong mechanism of catalysis. All together, these findings provide structural insights into the catalytic mechanism of dithiol Grxs.

DOI: 10.1021/bi401293s
PubMed: 24611845

Affiliations:

République populaire de Chine

Links toward previous steps (curation, corpus...)

to stream Main, to step Corpus: 000643
to stream Main, to step Curation: 000643

Le document en format XML

<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Structure-guided activity enhancement and catalytic mechanism of yeast grx8.</title>
<author><name sortKey="Tang, Yajun" sort="Tang, Yajun" uniqKey="Tang Y" first="Yajun" last="Tang">Yajun Tang</name>
<affiliation wicri:level="1"><nlm:affiliation>Hefei National Laboratory for Physical Sciences at the Microscale, School of Life Sciences, University of Science and Technology of China , Hefei, Anhui 230027, People's Republic of China.</nlm:affiliation>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>Hefei National Laboratory for Physical Sciences at the Microscale, School of Life Sciences, University of Science and Technology of China , Hefei, Anhui 230027</wicri:regionArea>
<wicri:noRegion>Anhui 230027</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Zhang, Jiahai" sort="Zhang, Jiahai" uniqKey="Zhang J" first="Jiahai" last="Zhang">Jiahai Zhang</name>
</author>
<author><name sortKey="Yu, Jiang" sort="Yu, Jiang" uniqKey="Yu J" first="Jiang" last="Yu">Jiang Yu</name>
</author>
<author><name sortKey="Xu, Ling" sort="Xu, Ling" uniqKey="Xu L" first="Ling" last="Xu">Ling Xu</name>
</author>
<author><name sortKey="Wu, Jihui" sort="Wu, Jihui" uniqKey="Wu J" first="Jihui" last="Wu">Jihui Wu</name>
</author>
<author><name sortKey="Zhou, Cong Zhao" sort="Zhou, Cong Zhao" uniqKey="Zhou C" first="Cong-Zhao" last="Zhou">Cong-Zhao Zhou</name>
</author>
<author><name sortKey="Shi, Yunyu" sort="Shi, Yunyu" uniqKey="Shi Y" first="Yunyu" last="Shi">Yunyu Shi</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="2014">2014</date>
<idno type="RBID">pubmed:24611845</idno>
<idno type="pmid">24611845</idno>
<idno type="doi">10.1021/bi401293s</idno>
<idno type="wicri:Area/Main/Corpus">000643</idno>
<idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000643</idno>
<idno type="wicri:Area/Main/Curation">000643</idno>
<idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000643</idno>
<idno type="wicri:Area/Main/Exploration">000643</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en">Structure-guided activity enhancement and catalytic mechanism of yeast grx8.</title>
<author><name sortKey="Tang, Yajun" sort="Tang, Yajun" uniqKey="Tang Y" first="Yajun" last="Tang">Yajun Tang</name>
<affiliation wicri:level="1"><nlm:affiliation>Hefei National Laboratory for Physical Sciences at the Microscale, School of Life Sciences, University of Science and Technology of China , Hefei, Anhui 230027, People's Republic of China.</nlm:affiliation>
<country xml:lang="fr">République populaire de Chine</country>
<wicri:regionArea>Hefei National Laboratory for Physical Sciences at the Microscale, School of Life Sciences, University of Science and Technology of China , Hefei, Anhui 230027</wicri:regionArea>
<wicri:noRegion>Anhui 230027</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Zhang, Jiahai" sort="Zhang, Jiahai" uniqKey="Zhang J" first="Jiahai" last="Zhang">Jiahai Zhang</name>
</author>
<author><name sortKey="Yu, Jiang" sort="Yu, Jiang" uniqKey="Yu J" first="Jiang" last="Yu">Jiang Yu</name>
</author>
<author><name sortKey="Xu, Ling" sort="Xu, Ling" uniqKey="Xu L" first="Ling" last="Xu">Ling Xu</name>
</author>
<author><name sortKey="Wu, Jihui" sort="Wu, Jihui" uniqKey="Wu J" first="Jihui" last="Wu">Jihui Wu</name>
</author>
<author><name sortKey="Zhou, Cong Zhao" sort="Zhou, Cong Zhao" uniqKey="Zhou C" first="Cong-Zhao" last="Zhou">Cong-Zhao Zhou</name>
</author>
<author><name sortKey="Shi, Yunyu" sort="Shi, Yunyu" uniqKey="Shi Y" first="Yunyu" last="Shi">Yunyu Shi</name>
</author>
</analytic>
<series><title level="j">Biochemistry</title>
<idno type="eISSN">1520-4995</idno>
<imprint><date when="2014" type="published">2014</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Biocatalysis (MeSH)</term>
<term>Enzyme Activation (MeSH)</term>
<term>Glutaredoxins (chemistry)</term>
<term>Glutaredoxins (isolation & purification)</term>
<term>Glutaredoxins (metabolism)</term>
<term>Kinetics (MeSH)</term>
<term>Nuclear Magnetic Resonance, Biomolecular (MeSH)</term>
<term>Protein Conformation (MeSH)</term>
<term>Recombinant Proteins (chemistry)</term>
<term>Recombinant Proteins (isolation & purification)</term>
<term>Recombinant Proteins (metabolism)</term>
<term>Saccharomyces cerevisiae (enzymology)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Activation enzymatique (MeSH)</term>
<term>Biocatalyse (MeSH)</term>
<term>Cinétique (MeSH)</term>
<term>Conformation des protéines (MeSH)</term>
<term>Glutarédoxines (composition chimique)</term>
<term>Glutarédoxines (isolement et purification)</term>
<term>Glutarédoxines (métabolisme)</term>
<term>Protéines recombinantes (composition chimique)</term>
<term>Protéines recombinantes (isolement et purification)</term>
<term>Protéines recombinantes (métabolisme)</term>
<term>Résonance magnétique nucléaire biomoléculaire (MeSH)</term>
<term>Saccharomyces cerevisiae (enzymologie)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Glutaredoxins</term>
<term>Recombinant Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Glutaredoxins</term>
<term>Recombinant Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Glutaredoxins</term>
<term>Recombinant Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Glutarédoxines</term>
<term>Protéines recombinantes</term>
</keywords>
<keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Saccharomyces cerevisiae</term>
</keywords>
<keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Saccharomyces cerevisiae</term>
</keywords>
<keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Glutarédoxines</term>
<term>Protéines recombinantes</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Glutarédoxines</term>
<term>Protéines recombinantes</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Biocatalysis</term>
<term>Enzyme Activation</term>
<term>Kinetics</term>
<term>Nuclear Magnetic Resonance, Biomolecular</term>
<term>Protein Conformation</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Activation enzymatique</term>
<term>Biocatalyse</term>
<term>Cinétique</term>
<term>Conformation des protéines</term>
<term>Résonance magnétique nucléaire biomoléculaire</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">Glutaredoxins (Grxs) are wide-spread oxidoreductases that are found in all kingdoms of life. The yeast Saccharomyces cerevisiae encodes eight Grxs, among which, Grx8 shares a sequence identity of 30 and 23% with typical dithiol Grx1 and Grx2, respectively, but it exhibits a much lower GSH-dependent oxidoreductase activity. To elucidate its catalytic mechanism, we solved the solution structure of Grx8, which displays a typical Grx fold. Structural analysis indicated that Grx8 possesses a negatively charged CXXC motif (Cys(33)-Pro(34)-Asp(35)-Cys(36)) and a GSH-recognition site, which are distinct from Grx1 and Grx2. Subsequent structure-guided site mutations revealed that the D35Y single mutant and N80T/L81V double mutant possess increased activity of 10- and 11-fold, respectively; moreover, the D35Y/N80T/L81V triple mutant has increased activity of up to 44-fold, which is comparable to that of canonical Grx. Biochemical analyses suggested that the increase in catalytic efficiency resulted from a decreased pKa value of catalytic cysteine Cys33 and/or enhancement of the putative GSH-recognition site. Moreover, NMR chemical shift perturbation analyses combined with GSH analogue inhibition assays enabled us to elucidate that wild-type Grx8 and all mutants adopt a ping-pong mechanism of catalysis. All together, these findings provide structural insights into the catalytic mechanism of dithiol Grxs. </div>
</front>
</TEI>
<pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">24611845</PMID>
<DateCompleted><Year>2014</Year>
<Month>06</Month>
<Day>23</Day>
</DateCompleted>
<DateRevised><Year>2014</Year>
<Month>04</Month>
<Day>09</Day>
</DateRevised>
<Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1520-4995</ISSN>
<JournalIssue CitedMedium="Internet"><Volume>53</Volume>
<Issue>13</Issue>
<PubDate><Year>2014</Year>
<Month>Apr</Month>
<Day>08</Day>
</PubDate>
</JournalIssue>
<Title>Biochemistry</Title>
<ISOAbbreviation>Biochemistry</ISOAbbreviation>
</Journal>
<ArticleTitle>Structure-guided activity enhancement and catalytic mechanism of yeast grx8.</ArticleTitle>
<Pagination><MedlinePgn>2185-96</MedlinePgn>
</Pagination>
<ELocationID EIdType="doi" ValidYN="Y">10.1021/bi401293s</ELocationID>
<Abstract><AbstractText>Glutaredoxins (Grxs) are wide-spread oxidoreductases that are found in all kingdoms of life. The yeast Saccharomyces cerevisiae encodes eight Grxs, among which, Grx8 shares a sequence identity of 30 and 23% with typical dithiol Grx1 and Grx2, respectively, but it exhibits a much lower GSH-dependent oxidoreductase activity. To elucidate its catalytic mechanism, we solved the solution structure of Grx8, which displays a typical Grx fold. Structural analysis indicated that Grx8 possesses a negatively charged CXXC motif (Cys(33)-Pro(34)-Asp(35)-Cys(36)) and a GSH-recognition site, which are distinct from Grx1 and Grx2. Subsequent structure-guided site mutations revealed that the D35Y single mutant and N80T/L81V double mutant possess increased activity of 10- and 11-fold, respectively; moreover, the D35Y/N80T/L81V triple mutant has increased activity of up to 44-fold, which is comparable to that of canonical Grx. Biochemical analyses suggested that the increase in catalytic efficiency resulted from a decreased pKa value of catalytic cysteine Cys33 and/or enhancement of the putative GSH-recognition site. Moreover, NMR chemical shift perturbation analyses combined with GSH analogue inhibition assays enabled us to elucidate that wild-type Grx8 and all mutants adopt a ping-pong mechanism of catalysis. All together, these findings provide structural insights into the catalytic mechanism of dithiol Grxs. </AbstractText>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Tang</LastName>
<ForeName>Yajun</ForeName>
<Initials>Y</Initials>
<AffiliationInfo><Affiliation>Hefei National Laboratory for Physical Sciences at the Microscale, School of Life Sciences, University of Science and Technology of China , Hefei, Anhui 230027, People's Republic of China.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Zhang</LastName>
<ForeName>Jiahai</ForeName>
<Initials>J</Initials>
</Author>
<Author ValidYN="Y"><LastName>Yu</LastName>
<ForeName>Jiang</ForeName>
<Initials>J</Initials>
</Author>
<Author ValidYN="Y"><LastName>Xu</LastName>
<ForeName>Ling</ForeName>
<Initials>L</Initials>
</Author>
<Author ValidYN="Y"><LastName>Wu</LastName>
<ForeName>Jihui</ForeName>
<Initials>J</Initials>
</Author>
<Author ValidYN="Y"><LastName>Zhou</LastName>
<ForeName>Cong-Zhao</ForeName>
<Initials>CZ</Initials>
</Author>
<Author ValidYN="Y"><LastName>Shi</LastName>
<ForeName>Yunyu</ForeName>
<Initials>Y</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>2014</Year>
<Month>03</Month>
<Day>25</Day>
</ArticleDate>
</Article>
<MedlineJournalInfo><Country>United States</Country>
<MedlineTA>Biochemistry</MedlineTA>
<NlmUniqueID>0370623</NlmUniqueID>
<ISSNLinking>0006-2960</ISSNLinking>
</MedlineJournalInfo>
<ChemicalList><Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="C000588987">GRX8 protein, S cerevisiae</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D054477">Glutaredoxins</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance>
</Chemical>
</ChemicalList>
<CitationSubset>IM</CitationSubset>
<MeshHeadingList><MeshHeading><DescriptorName UI="D055162" MajorTopicYN="Y">Biocatalysis</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D004789" MajorTopicYN="N">Enzyme Activation</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D054477" MajorTopicYN="N">Glutaredoxins</DescriptorName>
<QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName>
<QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D007700" MajorTopicYN="N">Kinetics</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D019906" MajorTopicYN="N">Nuclear Magnetic Resonance, Biomolecular</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D011487" MajorTopicYN="N">Protein Conformation</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName>
<QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName>
<QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D012441" MajorTopicYN="N">Saccharomyces cerevisiae</DescriptorName>
<QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName>
</MeshHeading>
</MeshHeadingList>
</MedlineCitation>
<PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2014</Year>
<Month>3</Month>
<Day>12</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
<PubMedPubDate PubStatus="pubmed"><Year>2014</Year>
<Month>3</Month>
<Day>13</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
<PubMedPubDate PubStatus="medline"><Year>2014</Year>
<Month>6</Month>
<Day>24</Day>
<Hour>6</Hour>
<Minute>0</Minute>
</PubMedPubDate>
</History>
<PublicationStatus>ppublish</PublicationStatus>
<ArticleIdList><ArticleId IdType="pubmed">24611845</ArticleId>
<ArticleId IdType="doi">10.1021/bi401293s</ArticleId>
</ArticleIdList>
</PubmedData>
</pubmed>
<affiliations><list><country><li>République populaire de Chine</li>
</country>
</list>
<tree><noCountry><name sortKey="Shi, Yunyu" sort="Shi, Yunyu" uniqKey="Shi Y" first="Yunyu" last="Shi">Yunyu Shi</name>
<name sortKey="Wu, Jihui" sort="Wu, Jihui" uniqKey="Wu J" first="Jihui" last="Wu">Jihui Wu</name>
<name sortKey="Xu, Ling" sort="Xu, Ling" uniqKey="Xu L" first="Ling" last="Xu">Ling Xu</name>
<name sortKey="Yu, Jiang" sort="Yu, Jiang" uniqKey="Yu J" first="Jiang" last="Yu">Jiang Yu</name>
<name sortKey="Zhang, Jiahai" sort="Zhang, Jiahai" uniqKey="Zhang J" first="Jiahai" last="Zhang">Jiahai Zhang</name>
<name sortKey="Zhou, Cong Zhao" sort="Zhou, Cong Zhao" uniqKey="Zhou C" first="Cong-Zhao" last="Zhou">Cong-Zhao Zhou</name>
</noCountry>
<country name="République populaire de Chine"><noRegion><name sortKey="Tang, Yajun" sort="Tang, Yajun" uniqKey="Tang Y" first="Yajun" last="Tang">Yajun Tang</name>
</noRegion>
</country>
</tree>
</affiliations>
</record>

Pour manipuler ce document sous Unix (Dilib)

EXPLOR_STEP=$WICRI_ROOT/Bois/explor/GlutaredoxinV1/Data/Main/Exploration

HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000598 | SxmlIndent | more

HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 000598 | SxmlIndent | more

Pour mettre un lien sur cette page dans le réseau Wicri

{{Explor lien
   |wiki=    Bois
   |area=    GlutaredoxinV1
   |flux=    Main
   |étape=   Exploration
   |type=    RBID
   |clé=     pubmed:24611845
   |texte=   Structure-guided activity enhancement and catalytic mechanism of yeast grx8.
}}

Pour générer des pages wiki

HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i   -Sk "pubmed:24611845" \
       | HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd   \
       | NlmPubMed2Wicri -a GlutaredoxinV1

This area was generated with Dilib version V0.6.37.
Data generation: Wed Nov 18 15:13:42 2020. Site generation: Wed Nov 18 15:16:12 2020

	Serveur d'exploration sur la glutarédoxine
	Attention, ce site est en cours de développement ! Attention, site généré par des moyens informatiques à partir de corpus bruts. Les informations ne sont donc pas validées.

Serveur d'exploration sur la glutarédoxine

Structure-guided activity enhancement and catalytic mechanism of yeast grx8.

Structure-guided activity enhancement and catalytic mechanism of yeast grx8.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki