Binding, internalization, and degradation of basic fibroblast growth factor in human microvascular endothelial cells
Identifieur interne : 003140 ( Main/Exploration ); précédent : 003139; suivant : 003141Binding, internalization, and degradation of basic fibroblast growth factor in human microvascular endothelial cells
Auteurs : A. Bikfalvi [France] ; E. Dupuy [France] ; A. L. Inyang [France] ; N. Fayein [France] ; G. Leseche [France] ; Y. Courtois [France] ; G. Tobelem [France]Source :
- Experimental Cell Research [ 0014-4827 ] ; 1989.
Descripteurs français
- KwdFr :
- MESH :
- métabolisme : Endothélium vasculaire, Facteurs de croissance fibroblastique, Membrane cellulaire.
- pharmacologie : Chloroquine.
- Cellules cultivées, Concentration en ions d'hydrogène, Humains, Masse moléculaire, Sites de fixation, Température.
English descriptors
- KwdEn :
- MESH :
- chemical , metabolism : Fibroblast Growth Factors.
- chemical , pharmacology : Chloroquine.
- metabolism : Cell Membrane, Endothelium, Vascular.
- Teeft :
- Acad, Baby hamster kidney cells, Bfgf, Bfgf binding, Binding Sites, Binding buffer, Binding data, Binding sites, Bovine capillary, Cell membrane, Cells, Cultured, Chloroquine, Chloroquine concentration, Cold binding buffer, Confluent home cells, Degradation, Degradation pattern, Degraded, Different fragments, Endothelial, Endothelial cells, Fresh binding medium, Growth factor, Hepes buffer, Home cells, Human microvascular, Human omental microvascular, Humans, Hydrogen-Ion Concentration, Incubation medium, Internalization, Ligand, Lowaffinity binding sites, Mitogenic signal, Molecular Weight, Murine lung capillary, Nacl, Nacl wash, Radioactivity, Same buffer, Several time intervals, Temperature, Time intervals, Total radioactivity, Unlabeled, Unlabeled bfgf, Unlabeled ligand, Vascular cells.
Abstract
Abstract: The binding, internalization, and degradation of basic fibroblast growth factor (bFGF) in human omental microvascular endothelial cells (HOME cells) were investigated. Binding studies of bFGF in human endothelial cells have not yet been reported. Basic FGF bound to HOME cells (KD of 42.0 ± 3.8 pM and 70,526 ± 6121 binding sites/cell for the high-affinity sites, KD of 0.933 ± 0.27 nM and 630,252 ± 172,459 sites/cell for low-affinity binding sites). The number of low-affinity binding sites was found to be variable. Washing the cells with 2 M phosphate-buffered saline removed completely 125I-bFGF bound to low-affinity binding sites but decreased also the high-affinity binding. The majority of the surface-bound 125I-bFGF was removed by washing the cells with acetic acid buffer at pH 3. At 37 °C, 30% of the cell-associated 125I-bFGF became resistant to the acidic wash after 90 min, suggesting that this fraction of bound 125I-bFGF was internalized. At this temperature, degradation of the internalized ligand was followed after 1 h by the appearance of three major bands of 15,000, 10,000, and 8,000 Da and was inhibited by chloroquine. These results demonstrated two classes of binding sites for bFGF in HOME cells; the number of high-affinity binding sites being larger than the number reported for bovine capillary endothelial cells. The intracellular processing of bFGF in HOME cells seems to be different from that of heparin binding growth factor-1 in murine lung capillary endothelial cells and of eye-derived growth factor-1 in Chinese hamster fibroblasts.
Url:
DOI: 10.1016/0014-4827(89)90183-3
Affiliations:
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Leseche</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Département de Chirurgie Vasculaire, Hôpital Beaujon, 92110 Clichy</wicri:regionArea><placeName><region type="region" nuts="2">Île-de-France</region><settlement type="city">Clichy</settlement></placeName></affiliation></author><author><name sortKey="Courtois, Y" sort="Courtois, Y" uniqKey="Courtois Y" first="Y." last="Courtois">Y. Courtois</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>INSERM U 118 and CNRS UA 630, 75016 Paris</wicri:regionArea><placeName><region type="region" nuts="2">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Tobelem, G" sort="Tobelem, G" uniqKey="Tobelem G" first="G." last="Tobelem">G. Tobelem</name><affiliation></affiliation><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>INSERM U 150 and CNRS UA 334, Hôpital Lariboisière, 75010 Paris</wicri:regionArea><placeName><region type="region" nuts="2">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">Experimental Cell Research</title><title level="j" type="abbrev">YEXCR</title><idno type="ISSN">0014-4827</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1989">1989</date><biblScope unit="volume">181</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="75">75</biblScope><biblScope unit="page" to="84">84</biblScope></imprint><idno type="ISSN">0014-4827</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0014-4827</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Binding Sites</term><term>Cell Membrane (metabolism)</term><term>Cells, Cultured</term><term>Chloroquine (pharmacology)</term><term>Endothelium, Vascular (metabolism)</term><term>Fibroblast Growth Factors (metabolism)</term><term>Humans</term><term>Hydrogen-Ion Concentration</term><term>Molecular Weight</term><term>Temperature</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Cellules cultivées</term><term>Chloroquine (pharmacologie)</term><term>Concentration en ions d'hydrogène</term><term>Endothélium vasculaire (métabolisme)</term><term>Facteurs de croissance fibroblastique (métabolisme)</term><term>Humains</term><term>Masse moléculaire</term><term>Membrane cellulaire (métabolisme)</term><term>Sites de fixation</term><term>Température</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Fibroblast Growth Factors</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Chloroquine</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Cell Membrane</term><term>Endothelium, Vascular</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Endothélium vasculaire</term><term>Facteurs de croissance fibroblastique</term><term>Membrane cellulaire</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Chloroquine</term></keywords><keywords scheme="Teeft" xml:lang="en"><term>Acad</term><term>Baby hamster kidney cells</term><term>Bfgf</term><term>Bfgf binding</term><term>Binding Sites</term><term>Binding buffer</term><term>Binding data</term><term>Binding sites</term><term>Bovine capillary</term><term>Cell membrane</term><term>Cells, Cultured</term><term>Chloroquine</term><term>Chloroquine concentration</term><term>Cold binding buffer</term><term>Confluent home cells</term><term>Degradation</term><term>Degradation pattern</term><term>Degraded</term><term>Different fragments</term><term>Endothelial</term><term>Endothelial cells</term><term>Fresh binding medium</term><term>Growth factor</term><term>Hepes buffer</term><term>Home cells</term><term>Human microvascular</term><term>Human omental microvascular</term><term>Humans</term><term>Hydrogen-Ion Concentration</term><term>Incubation medium</term><term>Internalization</term><term>Ligand</term><term>Lowaffinity binding sites</term><term>Mitogenic signal</term><term>Molecular Weight</term><term>Murine lung capillary</term><term>Nacl</term><term>Nacl wash</term><term>Radioactivity</term><term>Same buffer</term><term>Several time intervals</term><term>Temperature</term><term>Time intervals</term><term>Total radioactivity</term><term>Unlabeled</term><term>Unlabeled bfgf</term><term>Unlabeled ligand</term><term>Vascular cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Cellules cultivées</term><term>Concentration en ions d'hydrogène</term><term>Humains</term><term>Masse moléculaire</term><term>Sites de fixation</term><term>Température</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: The binding, internalization, and degradation of basic fibroblast growth factor (bFGF) in human omental microvascular endothelial cells (HOME cells) were investigated. Binding studies of bFGF in human endothelial cells have not yet been reported. Basic FGF bound to HOME cells (KD of 42.0 ± 3.8 pM and 70,526 ± 6121 binding sites/cell for the high-affinity sites, KD of 0.933 ± 0.27 nM and 630,252 ± 172,459 sites/cell for low-affinity binding sites). The number of low-affinity binding sites was found to be variable. Washing the cells with 2 M phosphate-buffered saline removed completely 125I-bFGF bound to low-affinity binding sites but decreased also the high-affinity binding. The majority of the surface-bound 125I-bFGF was removed by washing the cells with acetic acid buffer at pH 3. At 37 °C, 30% of the cell-associated 125I-bFGF became resistant to the acidic wash after 90 min, suggesting that this fraction of bound 125I-bFGF was internalized. At this temperature, degradation of the internalized ligand was followed after 1 h by the appearance of three major bands of 15,000, 10,000, and 8,000 Da and was inhibited by chloroquine. These results demonstrated two classes of binding sites for bFGF in HOME cells; the number of high-affinity binding sites being larger than the number reported for bovine capillary endothelial cells. The intracellular processing of bFGF in HOME cells seems to be different from that of heparin binding growth factor-1 in murine lung capillary endothelial cells and of eye-derived growth factor-1 in Chinese hamster fibroblasts.</div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Île-de-France</li></region><settlement><li>Clichy</li><li>Paris</li></settlement></list><tree><country name="France"><region name="Île-de-France"><name sortKey="Bikfalvi, A" sort="Bikfalvi, A" uniqKey="Bikfalvi A" first="A." last="Bikfalvi">A. Bikfalvi</name></region><name sortKey="Courtois, Y" sort="Courtois, Y" uniqKey="Courtois Y" first="Y." last="Courtois">Y. Courtois</name><name sortKey="Dupuy, E" sort="Dupuy, E" uniqKey="Dupuy E" first="E." last="Dupuy">E. Dupuy</name><name sortKey="Fayein, N" sort="Fayein, N" uniqKey="Fayein N" first="N." last="Fayein">N. Fayein</name><name sortKey="Inyang, A L" sort="Inyang, A L" uniqKey="Inyang A" first="A. L." last="Inyang">A. L. Inyang</name><name sortKey="Leseche, G" sort="Leseche, G" uniqKey="Leseche G" first="G." last="Leseche">G. Leseche</name><name sortKey="Tobelem, G" sort="Tobelem, G" uniqKey="Tobelem G" first="G." last="Tobelem">G. Tobelem</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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