Takeuchi Fusako: Protein-protein interaction of cytochrome b561 in chromaffin vesicle membranes studied by two-dimensional blue-native/sodium dodecyl sulfate gel electrophoresis and co-immunoprecipitation analysis. In: Acta biologica Szegediensis, (50) 1-2. pp. 83-87. (2006)
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We analyzed a protein-protein interaction in solubilized chromaffin vesicles using two-dimensional electrophoresis (1st, Blue-Native PAGE; 2nd, Tricine-SDS-PAGE). Cytochrome b561 band, which was verified by immunoblotting, was observed in the two-dimensional gel with an apparent molecular weight of ~100~400kDa. On the other hand, purified cytochrome b561 showed a monomeric band (28 kDa) in Blue-Native PAGE. These results indicated that cytochrome b561 interacts with other proteins in the chromaffin vesicles to form a large protein complex(es). To clarify the nature of the interaction, we performed co-immunoprecipitation experiments, where the solubilized membrane proteins were treated with immunopurified anti- b561 IgG antibodies followed by sedimentation with protein-A-Sepharose. We found that there were no other proteins co-sedimented with cytochrome b561. Since the immunopurified anti- b561 IgG antibodies bound specifically to the C-terminal hydrophilic portion of cytochrome b561 protein, we concluded that such binding of the IgG antibodies to the C-terminal portion might cause an inhibition of protein-protein interaction with other proteins in the solubilized state.
| Mű típusa: | Cikk, tanulmány, mű |
|---|---|
| Rovatcím: | Proceedings |
| Befoglaló folyóirat/kiadvány címe: | Acta biologica Szegediensis |
| Dátum: | 2006 |
| Kötet: | 50 |
| Szám: | 1-2 |
| ISSN: | 1588-385X |
| Oldalak: | pp. 83-87 |
| Nyelv: | angol |
| Konferencia neve: | International Conference on Membrane Redox Systems and Their Role in Biological Stress and Disease, 8., 2006, Szeged |
| Befoglaló mű URL: | http://acta.bibl.u-szeged.hu/39250/ |
| Kulcsszavak: | Természettudomány, Biológia |
| Megjegyzések: | Bibliogr.: 87. p.; Abstract |
| Feltöltés dátuma: | 2016. okt. 17. 09:24 |
| Utolsó módosítás: | 2021. ápr. 13. 10:56 |
| URI: | http://acta.bibl.u-szeged.hu/id/eprint/22776 |
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