Cloning of the rat CR3 alphaM (CD11b) subunit, expression and binding assay of recombinant isolated CD11b VA (A-domain) and ICAM-1 Ig modules.
Identifieur interne : 000016 ( Main/Exploration ); précédent : 000015; suivant : 000017Cloning of the rat CR3 alphaM (CD11b) subunit, expression and binding assay of recombinant isolated CD11b VA (A-domain) and ICAM-1 Ig modules.
Auteurs : K. Zerria [Tunisie] ; N. Bebbiche ; M R Barbouche ; K. Dellagi ; D M FathallahSource :
- Archives de l'Institut Pasteur de Tunis [ 0020-2509 ] ; 2002.
Descripteurs français
- KwdFr :
- ADN complémentaire (génétique), Adhérence cellulaire (génétique), Animaux (MeSH), Antigènes CD11b (génétique), Antigènes CD18 (génétique), Banque de gènes (MeSH), Chimiotaxie des leucocytes (génétique), Clonage moléculaire (méthodes), Données de séquences moléculaires (MeSH), Escherichia coli (génétique), Expression des gènes (génétique), Facteur de von Willebrand (génétique), Humains (MeSH), Molécule-1 d'adhérence intercellulaire (génétique), Protéines de fusion recombinantes (génétique), RT-PCR (MeSH), Rats (MeSH), Similitude de séquences (MeSH), Souris (MeSH), Séquence d'acides aminés (MeSH).
- MESH :
- génétique : ADN complémentaire, Adhérence cellulaire, Antigènes CD11b, Antigènes CD18, Chimiotaxie des leucocytes, Escherichia coli, Expression des gènes, Facteur de von Willebrand, Molécule-1 d'adhérence intercellulaire, Protéines de fusion recombinantes.
- méthodes : Clonage moléculaire.
- Animaux, Banque de gènes, Données de séquences moléculaires, Humains, RT-PCR, Rats, Similitude de séquences, Souris, Séquence d'acides aminés.
English descriptors
- KwdEn :
- Amino Acid Sequence (MeSH), Animals (MeSH), CD11b Antigen (genetics), CD18 Antigens (genetics), Cell Adhesion (genetics), Chemotaxis, Leukocyte (genetics), Cloning, Molecular (methods), DNA, Complementary (genetics), Escherichia coli (genetics), Gene Expression (genetics), Gene Library (MeSH), Humans (MeSH), Intercellular Adhesion Molecule-1 (genetics), Mice (MeSH), Molecular Sequence Data (MeSH), Rats (MeSH), Recombinant Fusion Proteins (genetics), Reverse Transcriptase Polymerase Chain Reaction (MeSH), Sequence Homology (MeSH), von Willebrand Factor (genetics).
- MESH :
- chemical , genetics : CD11b Antigen, CD18 Antigens, DNA, Complementary, Intercellular Adhesion Molecule-1, Recombinant Fusion Proteins, von Willebrand Factor.
- genetics : Cell Adhesion, Chemotaxis, Leukocyte, Escherichia coli, Gene Expression.
- methods : Cloning, Molecular.
- Amino Acid Sequence, Animals, Gene Library, Humans, Mice, Molecular Sequence Data, Rats, Reverse Transcriptase Polymerase Chain Reaction, Sequence Homology.
Abstract
The leukocyte beta2 integrin CR3 (CD11/CD18), is a surface heterodimeric glycoprotein that functions as a divalent cation-dependent adhesive complex. It mediates several important cell-substrate and cell-cell adhesive interactions among which the interaction with vascular endothelial cells that lead to leukocyte transmigration. We have isolated cDNA clones-coding for the rat complement receptor type 3 (CR3) alphaM subunit (CD11b) from a cDNA library. The cDNA sequence showed respectively 89.4% and 74.6% homology with its mouse and human counterpart. We have expressed the sequence coding for the VA module or Von Willebrand type domain (A-domain) and produced it in E. coli as a soluble recombinant fusion protein with GST. Simultaneously, we have cloned DNA fragments specific to the rat ICAM-1 domain 1 and domain 3 and expressed each clone in E. coli as recombinant soluble (rs) fusion proteins with GST. Recombinant CD11b A-domain was released from the fusion protein by thrombin cut. Purified ICAM-1 fusion peptides and CD11b A-domain were used to develop a direct binding assay that showed a specific binding between the rat ICAM-1 Ig like domain 3 and CD11b A-domain. These data demonstrate that the IgSF modules can be produced as a soluble recombinant fusion protein and used to study direct binding to the VA module displayed by members of the integrin superfamily.
PubMed: 15072240
Affiliations:
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Le document en format XML
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Bebbiche</name></author><author><name sortKey="Barbouche, M R" sort="Barbouche, M R" uniqKey="Barbouche M" first="M R" last="Barbouche">M R Barbouche</name></author><author><name sortKey="Dellagi, K" sort="Dellagi, K" uniqKey="Dellagi K" first="K" last="Dellagi">K. Dellagi</name></author><author><name sortKey="Fathallah, D M" sort="Fathallah, D M" uniqKey="Fathallah D" first="D M" last="Fathallah">D M Fathallah</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2002">2002</date><idno type="RBID">pubmed:15072240</idno><idno type="pmid">15072240</idno><idno type="wicri:Area/PubMed/Corpus">000808</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000808</idno><idno type="wicri:Area/PubMed/Curation">000804</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">000804</idno><idno type="wicri:Area/PubMed/Checkpoint">000813</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">000813</idno><idno type="wicri:Area/Main/Merge">000016</idno><idno type="wicri:Area/Main/Curation">000016</idno><idno type="wicri:Area/Main/Exploration">000016</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Cloning of the rat CR3 alphaM (CD11b) subunit, expression and binding assay of recombinant isolated CD11b VA (A-domain) and ICAM-1 Ig modules.</title><author><name sortKey="Zerria, K" sort="Zerria, K" uniqKey="Zerria K" first="K" last="Zerria">K. Zerria</name><affiliation wicri:level="1"><nlm:affiliation>Molecular Biotechnology Group, Immunology Laboratory, Institut Pasteur de Tunis, Tunisia.</nlm:affiliation><country xml:lang="fr">Tunisie</country><wicri:regionArea>Molecular Biotechnology Group, Immunology Laboratory, Institut Pasteur de Tunis</wicri:regionArea><wicri:noRegion>Institut Pasteur de Tunis</wicri:noRegion></affiliation></author><author><name sortKey="Bebbiche, N" sort="Bebbiche, N" uniqKey="Bebbiche N" first="N" last="Bebbiche">N. Bebbiche</name></author><author><name sortKey="Barbouche, M R" sort="Barbouche, M R" uniqKey="Barbouche M" first="M R" last="Barbouche">M R Barbouche</name></author><author><name sortKey="Dellagi, K" sort="Dellagi, K" uniqKey="Dellagi K" first="K" last="Dellagi">K. Dellagi</name></author><author><name sortKey="Fathallah, D M" sort="Fathallah, D M" uniqKey="Fathallah D" first="D M" last="Fathallah">D M Fathallah</name></author></analytic><series><title level="j">Archives de l'Institut Pasteur de Tunis</title><idno type="ISSN">0020-2509</idno><imprint><date when="2002" type="published">2002</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence (MeSH)</term><term>Animals (MeSH)</term><term>CD11b Antigen (genetics)</term><term>CD18 Antigens (genetics)</term><term>Cell Adhesion (genetics)</term><term>Chemotaxis, Leukocyte (genetics)</term><term>Cloning, Molecular (methods)</term><term>DNA, Complementary (genetics)</term><term>Escherichia coli (genetics)</term><term>Gene Expression (genetics)</term><term>Gene Library (MeSH)</term><term>Humans (MeSH)</term><term>Intercellular Adhesion Molecule-1 (genetics)</term><term>Mice (MeSH)</term><term>Molecular Sequence Data (MeSH)</term><term>Rats (MeSH)</term><term>Recombinant Fusion Proteins (genetics)</term><term>Reverse Transcriptase Polymerase Chain Reaction (MeSH)</term><term>Sequence Homology (MeSH)</term><term>von Willebrand Factor (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN complémentaire (génétique)</term><term>Adhérence cellulaire (génétique)</term><term>Animaux (MeSH)</term><term>Antigènes CD11b (génétique)</term><term>Antigènes CD18 (génétique)</term><term>Banque de gènes (MeSH)</term><term>Chimiotaxie des leucocytes (génétique)</term><term>Clonage moléculaire (méthodes)</term><term>Données de séquences moléculaires (MeSH)</term><term>Escherichia coli (génétique)</term><term>Expression des gènes (génétique)</term><term>Facteur de von Willebrand (génétique)</term><term>Humains (MeSH)</term><term>Molécule-1 d'adhérence intercellulaire (génétique)</term><term>Protéines de fusion recombinantes (génétique)</term><term>RT-PCR (MeSH)</term><term>Rats (MeSH)</term><term>Similitude de séquences (MeSH)</term><term>Souris (MeSH)</term><term>Séquence d'acides aminés (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>CD11b Antigen</term><term>CD18 Antigens</term><term>DNA, Complementary</term><term>Intercellular Adhesion Molecule-1</term><term>Recombinant Fusion Proteins</term><term>von Willebrand Factor</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Cell Adhesion</term><term>Chemotaxis, Leukocyte</term><term>Escherichia coli</term><term>Gene Expression</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN complémentaire</term><term>Adhérence cellulaire</term><term>Antigènes CD11b</term><term>Antigènes CD18</term><term>Chimiotaxie des leucocytes</term><term>Escherichia coli</term><term>Expression des gènes</term><term>Facteur de von Willebrand</term><term>Molécule-1 d'adhérence intercellulaire</term><term>Protéines de fusion recombinantes</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Cloning, Molecular</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Clonage moléculaire</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Gene Library</term><term>Humans</term><term>Mice</term><term>Molecular Sequence Data</term><term>Rats</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Sequence Homology</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Banque de gènes</term><term>Données de séquences moléculaires</term><term>Humains</term><term>RT-PCR</term><term>Rats</term><term>Similitude de séquences</term><term>Souris</term><term>Séquence d'acides aminés</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The leukocyte beta2 integrin CR3 (CD11/CD18), is a surface heterodimeric glycoprotein that functions as a divalent cation-dependent adhesive complex. It mediates several important cell-substrate and cell-cell adhesive interactions among which the interaction with vascular endothelial cells that lead to leukocyte transmigration. We have isolated cDNA clones-coding for the rat complement receptor type 3 (CR3) alphaM subunit (CD11b) from a cDNA library. The cDNA sequence showed respectively 89.4% and 74.6% homology with its mouse and human counterpart. We have expressed the sequence coding for the VA module or Von Willebrand type domain (A-domain) and produced it in E. coli as a soluble recombinant fusion protein with GST. Simultaneously, we have cloned DNA fragments specific to the rat ICAM-1 domain 1 and domain 3 and expressed each clone in E. coli as recombinant soluble (rs) fusion proteins with GST. Recombinant CD11b A-domain was released from the fusion protein by thrombin cut. Purified ICAM-1 fusion peptides and CD11b A-domain were used to develop a direct binding assay that showed a specific binding between the rat ICAM-1 Ig like domain 3 and CD11b A-domain. These data demonstrate that the IgSF modules can be produced as a soluble recombinant fusion protein and used to study direct binding to the VA module displayed by members of the integrin superfamily.</div></front></TEI><affiliations><list><country><li>Tunisie</li></country></list><tree><noCountry><name sortKey="Barbouche, M R" sort="Barbouche, M R" uniqKey="Barbouche M" first="M R" last="Barbouche">M R Barbouche</name><name sortKey="Bebbiche, N" sort="Bebbiche, N" uniqKey="Bebbiche N" first="N" last="Bebbiche">N. Bebbiche</name><name sortKey="Dellagi, K" sort="Dellagi, K" uniqKey="Dellagi K" first="K" last="Dellagi">K. Dellagi</name><name sortKey="Fathallah, D M" sort="Fathallah, D M" uniqKey="Fathallah D" first="D M" last="Fathallah">D M Fathallah</name></noCountry><country name="Tunisie"><noRegion><name sortKey="Zerria, K" sort="Zerria, K" uniqKey="Zerria K" first="K" last="Zerria">K. Zerria</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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