کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5132065 | 1378790 | 2017 | 11 صفحه PDF | دانلود رایگان |
- Bcl-2Î21 interacts with and partially inhibits SERCA3b isoform.
- Bcl-2 interacts with and partially inhibits SERCA3b in cell culture.
- Both exogenous Bcl-2Î21 and endogenous Bcl-2 could form a stable immunocomplex with SERCA3b.
- Overexpression of Bcl-2 reduced fluorescein isothiocyanate (FITC) labeling of SERCA3b.
An interaction of Bcl-2 with SERCA had been documented in vitro using the SERCA1a isoform isolated from rat skeletal muscle [Dremina, E. S., Sharov, V. S., Kumar, K., Azidi, A., Michaelis, E. K., Schöneich, C. (2004) Biochem. J. 383 (361-370)]. Here, we demonstrate the interaction of Bcl-2 with the SERCA3b isoform both in vitro and in cell culture. In vitro, the interaction of Bcl-2 with SERCA3b was studied using Bcl-2â21, a truncated form of human Bcl-2, and microsomes isolated from SERCA3b-overexpressing HEK-293 cells. For these experiments, SERCA3b was quantified by a combination of amino acid analysis and Western blotting. We observed that Bcl-2â21 both inactivates SERCA3b and co-immunoprecipitates with SERCA3b. The incubation with Bcl-2â21 changes the distribution of SERCA3b during sucrose density gradient centrifugation, likely as the result of Bcl-2â21-induced conformational change of SERCA3b. When SERCA3b-overexpressing HEK-293 cells were co-transfected with Bcl-2, Bcl-2-dependent SERCA3b inactivation was observed. In these cells, Bcl-2 interaction with SERCA3b was demonstrated by co-immunoprecipitation. Furthermore, overexpression of Bcl-2 reduced fluorescein isothiocyanate (FITC) labeling of SERCA3b. Together, our data provide evidence for the interaction of Bcl-2 with SERCA3b in vitro and in cell culture, and for Bcl-2-dependent conformational and functional changes of SERCA3b.
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1865, Issue 1, January 2017, Pages 121-131