Conformational changes and in vitro core-formation modifications induced by site-directed mutagenesis of the specific N-terminus of pea seed ferritin.
Identifieur interne : 000324 ( France/Analysis ); précédent : 000323; suivant : 000325Conformational changes and in vitro core-formation modifications induced by site-directed mutagenesis of the specific N-terminus of pea seed ferritin.
Auteurs : O. Van Wuytswinkel [France] ; J F BriatSource :
- The Biochemical journal [ 0264-6021 ] ; 1995.
Descripteurs français
- KwdFr :
- Conformation des protéines, Données de séquences moléculaires, Dénaturation des protéines, Ferritines (), Ferritines (génétique), Fragments peptidiques (), Graines (), Mutagenèse dirigée, Pois, Protéines recombinantes (), Protéines végétales (), Relation structure-activité, Structure secondaire des protéines, Séquence d'acides aminés, Séquence nucléotidique, Thermodynamique, Urée.
- MESH :
- génétique : Ferritines.
- Conformation des protéines, Données de séquences moléculaires, Dénaturation des protéines, Ferritines, Fragments peptidiques, Graines, Mutagenèse dirigée, Pois, Protéines recombinantes, Protéines végétales, Relation structure-activité, Structure secondaire des protéines, Séquence d'acides aminés, Séquence nucléotidique, Thermodynamique, Urée.
English descriptors
- KwdEn :
- Amino Acid Sequence, Base Sequence, Ferritins (chemistry), Ferritins (genetics), Molecular Sequence Data, Mutagenesis, Site-Directed, Peas, Peptide Fragments (chemistry), Plant Proteins (chemistry), Protein Conformation, Protein Denaturation, Protein Structure, Secondary, Recombinant Proteins (chemistry), Seeds (chemistry), Structure-Activity Relationship, Thermodynamics, Urea.
- MESH :
- chemical , chemistry : Ferritins, Peptide Fragments, Plant Proteins, Recombinant Proteins.
- chemical , genetics : Ferritins.
- chemistry : Seeds.
- Amino Acid Sequence, Base Sequence, Molecular Sequence Data, Mutagenesis, Site-Directed, Peas, Protein Conformation, Protein Denaturation, Protein Structure, Secondary, Structure-Activity Relationship, Thermodynamics, Urea.
Abstract
Plant ferritin has a three-dimensional structure predicted to be very similar to that of animal ferritin. It has, however, an additional specific sequence of 24 amino acids at its N-terminus named extension peptide (EP). In order to determine precisely the interactions between EP and other domains of the pea seed ferritin subunit, three point mutations were performed. The mutated residues were chosen by three-dimensional computer modelling of the pea seed ferritin subunit structure [Lobréaux, Yewdall, Briat and Harrison (1992) Biochem. J. 228, 931-939]. The mutant recombinant proteins were expressed in Escherichia coli and purified to homogeneity; all the mutants were found to be assembled as 24-mers. When Ala-13 was replaced by His, as in mammalian ferritins, ferroxidase activity was significantly reduced. Moreover, in vitro iron-core formation in Pro-X-->Ala, Lys-R-->Glu and Ala-13-->His mutants was increased after denaturation by urea followed by renaturation; this was also observed with the EP deletion mutant (r delta TP/EP). The recombinant ferritins were also analysed using tryptophan fluorescence spectra. The r delta TP/EP, Pro-X-->Ala and Lys-R-->Glu mutants were found to be more susceptible to denaturation by urea than the native r delta TP pea seed ferritin.
DOI: 10.1042/bj3050959
PubMed: 7848297
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: 002833
- to stream PubMed, to step Curation: 002833
- to stream PubMed, to step Checkpoint: 002693
- to stream Ncbi, to step Merge: 002933
- to stream Ncbi, to step Curation: 002933
- to stream Ncbi, to step Checkpoint: 002933
- to stream Main, to step Merge: 004113
- to stream Main, to step Curation: 004053
- to stream Main, to step Exploration: 004053
- to stream France, to step Extraction: 000324
Links to Exploration step
pubmed:7848297Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Conformational changes and in vitro core-formation modifications induced by site-directed mutagenesis of the specific N-terminus of pea seed ferritin.</title><author><name sortKey="Van Wuytswinkel, O" sort="Van Wuytswinkel, O" uniqKey="Van Wuytswinkel O" first="O" last="Van Wuytswinkel">O. Van Wuytswinkel</name><affiliation wicri:level="3"><nlm:affiliation>Laboratoire de Biologie Moléculaire Végétale, Centre National de la Recherche Scientifique (Unité de Recherche 1178), Grenoble, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire de Biologie Moléculaire Végétale, Centre National de la Recherche Scientifique (Unité de Recherche 1178), Grenoble</wicri:regionArea><placeName><region type="region">Auvergne-Rhône-Alpes</region><region type="old region">Rhône-Alpes</region><settlement type="city">Grenoble</settlement></placeName></affiliation></author><author><name sortKey="Briat, J F" sort="Briat, J F" uniqKey="Briat J" first="J F" last="Briat">J F Briat</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1995">1995</date><idno type="RBID">pubmed:7848297</idno><idno type="pmid">7848297</idno><idno type="doi">10.1042/bj3050959</idno><idno type="wicri:Area/PubMed/Corpus">002833</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002833</idno><idno type="wicri:Area/PubMed/Curation">002833</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002833</idno><idno type="wicri:Area/PubMed/Checkpoint">002693</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002693</idno><idno type="wicri:Area/Ncbi/Merge">002933</idno><idno type="wicri:Area/Ncbi/Curation">002933</idno><idno type="wicri:Area/Ncbi/Checkpoint">002933</idno><idno type="wicri:doubleKey">0264-6021:1995:Van Wuytswinkel O:conformational:changes:and</idno><idno type="wicri:Area/Main/Merge">004113</idno><idno type="wicri:Area/Main/Curation">004053</idno><idno type="wicri:Area/Main/Exploration">004053</idno><idno type="wicri:Area/France/Extraction">000324</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Conformational changes and in vitro core-formation modifications induced by site-directed mutagenesis of the specific N-terminus of pea seed ferritin.</title><author><name sortKey="Van Wuytswinkel, O" sort="Van Wuytswinkel, O" uniqKey="Van Wuytswinkel O" first="O" last="Van Wuytswinkel">O. Van Wuytswinkel</name><affiliation wicri:level="3"><nlm:affiliation>Laboratoire de Biologie Moléculaire Végétale, Centre National de la Recherche Scientifique (Unité de Recherche 1178), Grenoble, France.</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire de Biologie Moléculaire Végétale, Centre National de la Recherche Scientifique (Unité de Recherche 1178), Grenoble</wicri:regionArea><placeName><region type="region">Auvergne-Rhône-Alpes</region><region type="old region">Rhône-Alpes</region><settlement type="city">Grenoble</settlement></placeName></affiliation></author><author><name sortKey="Briat, J F" sort="Briat, J F" uniqKey="Briat J" first="J F" last="Briat">J F Briat</name></author></analytic><series><title level="j">The Biochemical journal</title><idno type="ISSN">0264-6021</idno><imprint><date when="1995" type="published">1995</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Base Sequence</term><term>Ferritins (chemistry)</term><term>Ferritins (genetics)</term><term>Molecular Sequence Data</term><term>Mutagenesis, Site-Directed</term><term>Peas</term><term>Peptide Fragments (chemistry)</term><term>Plant Proteins (chemistry)</term><term>Protein Conformation</term><term>Protein Denaturation</term><term>Protein Structure, Secondary</term><term>Recombinant Proteins (chemistry)</term><term>Seeds (chemistry)</term><term>Structure-Activity Relationship</term><term>Thermodynamics</term><term>Urea</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Conformation des protéines</term><term>Données de séquences moléculaires</term><term>Dénaturation des protéines</term><term>Ferritines ()</term><term>Ferritines (génétique)</term><term>Fragments peptidiques ()</term><term>Graines ()</term><term>Mutagenèse dirigée</term><term>Pois</term><term>Protéines recombinantes ()</term><term>Protéines végétales ()</term><term>Relation structure-activité</term><term>Structure secondaire des protéines</term><term>Séquence d'acides aminés</term><term>Séquence nucléotidique</term><term>Thermodynamique</term><term>Urée</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Ferritins</term><term>Peptide Fragments</term><term>Plant Proteins</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Ferritins</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Seeds</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Ferritines</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Base Sequence</term><term>Molecular Sequence Data</term><term>Mutagenesis, Site-Directed</term><term>Peas</term><term>Protein Conformation</term><term>Protein Denaturation</term><term>Protein Structure, Secondary</term><term>Structure-Activity Relationship</term><term>Thermodynamics</term><term>Urea</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Conformation des protéines</term><term>Données de séquences moléculaires</term><term>Dénaturation des protéines</term><term>Ferritines</term><term>Fragments peptidiques</term><term>Graines</term><term>Mutagenèse dirigée</term><term>Pois</term><term>Protéines recombinantes</term><term>Protéines végétales</term><term>Relation structure-activité</term><term>Structure secondaire des protéines</term><term>Séquence d'acides aminés</term><term>Séquence nucléotidique</term><term>Thermodynamique</term><term>Urée</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Plant ferritin has a three-dimensional structure predicted to be very similar to that of animal ferritin. It has, however, an additional specific sequence of 24 amino acids at its N-terminus named extension peptide (EP). In order to determine precisely the interactions between EP and other domains of the pea seed ferritin subunit, three point mutations were performed. The mutated residues were chosen by three-dimensional computer modelling of the pea seed ferritin subunit structure [Lobréaux, Yewdall, Briat and Harrison (1992) Biochem. J. 228, 931-939]. The mutant recombinant proteins were expressed in Escherichia coli and purified to homogeneity; all the mutants were found to be assembled as 24-mers. When Ala-13 was replaced by His, as in mammalian ferritins, ferroxidase activity was significantly reduced. Moreover, in vitro iron-core formation in Pro-X-->Ala, Lys-R-->Glu and Ala-13-->His mutants was increased after denaturation by urea followed by renaturation; this was also observed with the EP deletion mutant (r delta TP/EP). The recombinant ferritins were also analysed using tryptophan fluorescence spectra. The r delta TP/EP, Pro-X-->Ala and Lys-R-->Glu mutants were found to be more susceptible to denaturation by urea than the native r delta TP pea seed ferritin.</div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Auvergne-Rhône-Alpes</li><li>Rhône-Alpes</li></region><settlement><li>Grenoble</li></settlement></list><tree><noCountry><name sortKey="Briat, J F" sort="Briat, J F" uniqKey="Briat J" first="J F" last="Briat">J F Briat</name></noCountry><country name="France"><region name="Auvergne-Rhône-Alpes"><name sortKey="Van Wuytswinkel, O" sort="Van Wuytswinkel, O" uniqKey="Van Wuytswinkel O" first="O" last="Van Wuytswinkel">O. Van Wuytswinkel</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/France/Analysis
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000324 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/France/Analysis/biblio.hfd -nk 000324 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= France
|étape= Analysis
|type= RBID
|clé= pubmed:7848297
|texte= Conformational changes and in vitro core-formation modifications induced by site-directed mutagenesis of the specific N-terminus of pea seed ferritin.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/France/Analysis/RBID.i -Sk "pubmed:7848297" \
| HfdSelect -Kh $EXPLOR_AREA/Data/France/Analysis/biblio.hfd \
| NlmPubMed2Wicri -a MersV1
| This area was generated with Dilib version V0.6.33. |  |