A member of the selectin family (gmp-140/padgem) is expressed on thrombin-stimulated rat platelets in vitro
Identifieur interne : 001538 ( Main/Exploration ); précédent : 001537; suivant : 001539A member of the selectin family (gmp-140/padgem) is expressed on thrombin-stimulated rat platelets in vitro
Auteurs : P. D Winocour [France] ; E. Chignier [France] ; S. Parmentier [France] ; J. L Mcgregor [France]Source :
- Comparative Biochemistry and Physiology -- Part A: Physiology [ 0300-9629 ] ; 1991.
Abstract
1.1. Granule membrane protein (GMP-140) is an integral α-granule membrane glycoprotein, expressed on the surface of human platelets following degranulation, and is pan of a new family of adhesion molecules (selectins) related to the endothelial leukocyte adhesion molecule (ELAM-1) and to the lymphocyte homing receptors in man (Leu-8/TQ1) and in mouse (gp90MEL-14).2.2. The cross-reactivity with rat platelets of the monoclonal antibodies (MAb), LYP20 and S12, directed against human GMP-140 was examined, with the purpose of assessing the homology of GMP-140 between human and rat platelets and of using positive MAbs to detect platelet activation in vivo in response to vascular disease in rats.3.3. By ELISA technique, LYP20 gave a greater OD reading with thrombin-stimulated rat platelets than with resting platelets.4.4. 125I-LYP20 bound significantly more to thrombin-stimulated rat platelets (3875 ± 750 molecules/ platelet) than to resting platelets (645 ± 240 molecules/platelet, P < 0.01) with 50% maximum binding at 0.13 ± 0.02 μg/ml; 125I-S12 did not bind to rat platelets.5.5. By fluorescence-activated flow cytometry there were significantly more fluorescent thrombin-stimulated platelets (56 ± 7% of total), compared with resting platelets (8 ± 1% of total, P < 0.001).6.6. Western blots of rat platelet lysates showed that LYP20 bound to a single band identified, under non-reducing conditions, as having the same apparent Mr as GMP-140.7.7. LYP20 immunoprecipitated a protein which became radiolabelled on the surface of thrombin-activated rat platelets; S12 did not recognize any protein.8.8. It appears that LYP20 and S12 are directed against different epitopes on the GMP-140 molecule and that the LYP20 epitope, but not the S12 epitope, is preserved in GMP-140 in rat platelets.9.9. LYP20 can, therefore, be used as a marker of GMP-140 expression on activated platelets in vivo in rats.
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DOI: 10.1016/0300-9629(92)90133-B
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L Mcgregor</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>INSERM Unit 331, Faculté de Médecine, Alexis Carrel, Institut Pasteur, F-69373 Lyon</wicri:regionArea><placeName><region type="region" nuts="2">Auvergne-Rhône-Alpes</region><region type="old region" nuts="2">Rhône-Alpes</region><settlement type="city">Lyon</settlement></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">Comparative Biochemistry and Physiology -- Part A: Physiology</title><title level="j" type="abbrev">CBAOLD</title><idno type="ISSN">0300-9629</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1991">1991</date><biblScope unit="volume">102</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="265">265</biblScope><biblScope unit="page" to="271">271</biblScope></imprint><idno type="ISSN">0300-9629</idno></series><idno type="istex">B9C49E9F497B93C8C60A3E28D81EBB3BC7CEDBC7</idno><idno type="DOI">10.1016/0300-9629(92)90133-B</idno><idno type="PII">0300-9629(92)90133-B</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0300-9629</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">1.1. Granule membrane protein (GMP-140) is an integral α-granule membrane glycoprotein, expressed on the surface of human platelets following degranulation, and is pan of a new family of adhesion molecules (selectins) related to the endothelial leukocyte adhesion molecule (ELAM-1) and to the lymphocyte homing receptors in man (Leu-8/TQ1) and in mouse (gp90MEL-14).2.2. The cross-reactivity with rat platelets of the monoclonal antibodies (MAb), LYP20 and S12, directed against human GMP-140 was examined, with the purpose of assessing the homology of GMP-140 between human and rat platelets and of using positive MAbs to detect platelet activation in vivo in response to vascular disease in rats.3.3. By ELISA technique, LYP20 gave a greater OD reading with thrombin-stimulated rat platelets than with resting platelets.4.4. 125I-LYP20 bound significantly more to thrombin-stimulated rat platelets (3875 ± 750 molecules/ platelet) than to resting platelets (645 ± 240 molecules/platelet, P < 0.01) with 50% maximum binding at 0.13 ± 0.02 μg/ml; 125I-S12 did not bind to rat platelets.5.5. By fluorescence-activated flow cytometry there were significantly more fluorescent thrombin-stimulated platelets (56 ± 7% of total), compared with resting platelets (8 ± 1% of total, P < 0.001).6.6. Western blots of rat platelet lysates showed that LYP20 bound to a single band identified, under non-reducing conditions, as having the same apparent Mr as GMP-140.7.7. LYP20 immunoprecipitated a protein which became radiolabelled on the surface of thrombin-activated rat platelets; S12 did not recognize any protein.8.8. It appears that LYP20 and S12 are directed against different epitopes on the GMP-140 molecule and that the LYP20 epitope, but not the S12 epitope, is preserved in GMP-140 in rat platelets.9.9. LYP20 can, therefore, be used as a marker of GMP-140 expression on activated platelets in vivo in rats.</div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Auvergne-Rhône-Alpes</li><li>Rhône-Alpes</li></region><settlement><li>Lyon</li></settlement></list><tree><country name="France"><region name="Auvergne-Rhône-Alpes"><name sortKey="Winocour, P D" sort="Winocour, P D" uniqKey="Winocour P" first="P. D" last="Winocour">P. D Winocour</name></region><name sortKey="Chignier, E" sort="Chignier, E" uniqKey="Chignier E" first="E" last="Chignier">E. Chignier</name><name sortKey="Mcgregor, J L" sort="Mcgregor, J L" uniqKey="Mcgregor J" first="J. L" last="Mcgregor">J. L Mcgregor</name><name sortKey="Parmentier, S" sort="Parmentier, S" uniqKey="Parmentier S" first="S" last="Parmentier">S. Parmentier</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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