Self-association of a small heat shock protein.
Identifieur interne : 002E81 ( Main/Exploration ); précédent : 002E80; suivant : 002E82Self-association of a small heat shock protein.
Auteurs : Barbara Lelj-Garolla [Canada] ; A Grant MaukSource :
- Journal of molecular biology [ 0022-2836 ] ; 2005.
Descripteurs français
- KwdFr :
- MESH :
- génétique : Protéines du choc thermique.
- métabolisme : Protéines du choc thermique.
- Délétion de séquence, Humains, Mutagenèse dirigée, Ultracentrifugation.
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : Heat-Shock Proteins.
- chemical , metabolism : Heat-Shock Proteins.
- Humans, Mutagenesis, Site-Directed, Sequence Deletion, Ultracentrifugation.
Abstract
Human Hsp27 oligomerizes in vivo in a phosphorylation-dependent manner that regulates the functional activity of the protein. We have studied the self-association of wild-type Hsp27 by both sedimentation velocity and sedimentation equilibrium analysis and established that the protein forms an equilibrium mixture of monomers/dimers, tetramers, 12-mers and 16-mers (20 mM Tris-HCl (pH 8.4), 100 mM NaCl, 20 degrees C). Corresponding analysis of the S15D/S78D/S82D triple variant, which is believed to mimic the behavior of phosphorylated Hsp27, establishes that this form of the protein forms primarily monomers and dimers but also forms a small fraction of very large oligomers. Variants in which critical N-terminal sequences have been deleted exhibit oligomerization behavior that is intermediate between that of the triple variant and the wild-type protein. On the other hand a C-terminal sequence deletion variant forms larger oligomers than does the wild-type protein, but also exhibits a greater fraction of smaller oligomers. Notably, the presence of an N-terminal His6-tag induces formation of much larger oligomers than observed for any other form of the protein. The results of this work establish that the wild-type protein forms smaller oligomers than previously believed, define the roles played by various structural domains in Hsp27 oligomerization, and provide improved molecular probes with better-defined properties for the design of future experiments.
DOI: 10.1016/j.jmb.2004.10.056
PubMed: 15581903
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: 002354
- to stream PubMed, to step Curation: 002354
- to stream PubMed, to step Checkpoint: 002179
- to stream Ncbi, to step Merge: 000323
- to stream Ncbi, to step Curation: 000323
- to stream Ncbi, to step Checkpoint: 000323
- to stream Main, to step Merge: 002F11
- to stream Main, to step Curation: 002E81
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Self-association of a small heat shock protein.</title><author><name sortKey="Lelj Garolla, Barbara" sort="Lelj Garolla, Barbara" uniqKey="Lelj Garolla B" first="Barbara" last="Lelj-Garolla">Barbara Lelj-Garolla</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, V6T 1Z3, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, V6T 1Z3</wicri:regionArea><wicri:noRegion>V6T 1Z3</wicri:noRegion></affiliation></author><author><name sortKey="Mauk, A Grant" sort="Mauk, A Grant" uniqKey="Mauk A" first="A Grant" last="Mauk">A Grant Mauk</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:15581903</idno><idno type="pmid">15581903</idno><idno type="doi">10.1016/j.jmb.2004.10.056</idno><idno type="wicri:Area/PubMed/Corpus">002354</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002354</idno><idno type="wicri:Area/PubMed/Curation">002354</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002354</idno><idno type="wicri:Area/PubMed/Checkpoint">002179</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002179</idno><idno type="wicri:Area/Ncbi/Merge">000323</idno><idno type="wicri:Area/Ncbi/Curation">000323</idno><idno type="wicri:Area/Ncbi/Checkpoint">000323</idno><idno type="wicri:doubleKey">0022-2836:2005:Lelj Garolla B:self:association:of</idno><idno type="wicri:Area/Main/Merge">002F11</idno><idno type="wicri:Area/Main/Curation">002E81</idno><idno type="wicri:Area/Main/Exploration">002E81</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Self-association of a small heat shock protein.</title><author><name sortKey="Lelj Garolla, Barbara" sort="Lelj Garolla, Barbara" uniqKey="Lelj Garolla B" first="Barbara" last="Lelj-Garolla">Barbara Lelj-Garolla</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, V6T 1Z3, Canada.</nlm:affiliation><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, V6T 1Z3</wicri:regionArea><wicri:noRegion>V6T 1Z3</wicri:noRegion></affiliation></author><author><name sortKey="Mauk, A Grant" sort="Mauk, A Grant" uniqKey="Mauk A" first="A Grant" last="Mauk">A Grant Mauk</name></author></analytic><series><title level="j">Journal of molecular biology</title><idno type="ISSN">0022-2836</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Heat-Shock Proteins (genetics)</term><term>Heat-Shock Proteins (metabolism)</term><term>Humans</term><term>Mutagenesis, Site-Directed</term><term>Sequence Deletion</term><term>Ultracentrifugation</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Délétion de séquence</term><term>Humains</term><term>Mutagenèse dirigée</term><term>Protéines du choc thermique (génétique)</term><term>Protéines du choc thermique (métabolisme)</term><term>Ultracentrifugation</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Heat-Shock Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Heat-Shock Proteins</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines du choc thermique</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Protéines du choc thermique</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Humans</term><term>Mutagenesis, Site-Directed</term><term>Sequence Deletion</term><term>Ultracentrifugation</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Délétion de séquence</term><term>Humains</term><term>Mutagenèse dirigée</term><term>Ultracentrifugation</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Human Hsp27 oligomerizes in vivo in a phosphorylation-dependent manner that regulates the functional activity of the protein. We have studied the self-association of wild-type Hsp27 by both sedimentation velocity and sedimentation equilibrium analysis and established that the protein forms an equilibrium mixture of monomers/dimers, tetramers, 12-mers and 16-mers (20 mM Tris-HCl (pH 8.4), 100 mM NaCl, 20 degrees C). Corresponding analysis of the S15D/S78D/S82D triple variant, which is believed to mimic the behavior of phosphorylated Hsp27, establishes that this form of the protein forms primarily monomers and dimers but also forms a small fraction of very large oligomers. Variants in which critical N-terminal sequences have been deleted exhibit oligomerization behavior that is intermediate between that of the triple variant and the wild-type protein. On the other hand a C-terminal sequence deletion variant forms larger oligomers than does the wild-type protein, but also exhibits a greater fraction of smaller oligomers. Notably, the presence of an N-terminal His6-tag induces formation of much larger oligomers than observed for any other form of the protein. The results of this work establish that the wild-type protein forms smaller oligomers than previously believed, define the roles played by various structural domains in Hsp27 oligomerization, and provide improved molecular probes with better-defined properties for the design of future experiments.</div></front></TEI><affiliations><list><country><li>Canada</li></country></list><tree><noCountry><name sortKey="Mauk, A Grant" sort="Mauk, A Grant" uniqKey="Mauk A" first="A Grant" last="Mauk">A Grant Mauk</name></noCountry><country name="Canada"><noRegion><name sortKey="Lelj Garolla, Barbara" sort="Lelj Garolla, Barbara" uniqKey="Lelj Garolla B" first="Barbara" last="Lelj-Garolla">Barbara Lelj-Garolla</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 002E81 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 002E81 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= pubmed:15581903
|texte= Self-association of a small heat shock protein.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:15581903" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a MersV1
| This area was generated with Dilib version V0.6.33. |  |