| Autor: |
Leopol'd AV; Department and Division of Medical Nanobiotechnologies, Medico-Biological Faculty, N. I. Pirogov Russian State Medical University, Russia. ahnavleopold@yandex.ru, Baklaushev VP, Korchagina AA, Shein SA, Grinenko NF, Pavlov KA, Ryabukhin IA, Chekhonin VP |
| Jazyk: |
English; Russian |
| Zdroj: |
Bulletin of experimental biology and medicine [Bull Exp Biol Med] 2012 Apr; Vol. 152 (6), pp. 707-11. |
| DOI: |
10.1007/s10517-012-1612-0 |
| Abstrakt: |
cDNA encoding VEGF and Ig-like extracellular domains 2-4 of VEGFR-1 (sFlt-1(2-4)) were cloned into prokaryotic expression vectors pET32a and pQE60. Recombinant proteins were purified (metal affinity chromatography) and renatured. Chemiluminescent study for the interaction of recombinant VEGF and sFlt-1(2-4) showed that biotinylated VEGF specifically binds to the polystyrene-immobilized receptor extracellular fragment. Biotinylated recombinant sFlt-1 interacts with immobilized VEGF. Analysis of the interaction of immobilized recombinant VEGFR-1 and VEGF with C6 glioma cells labeled with CFDA-SE (vital fluorescent dye) showed that recombinant VEGFR-1 also binds to native membrane-associated VEGF. Recombinant VEGF was shown to bind to specific receptors expressed on the surface of C6 glioma cells. Functional activity of these proteins was confirmed by ligand-receptor assay for VEGF and VEGFR-1 (sFlt-1) and quantitative chemiluminescent detection. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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