Model system for the analysis of cell surface expression of human ABCA1

Abstract Background The ABCA1 protein plays a pivotal role in reverse cholesterol transport, by mediating the generation of HDL particles and removing cellular cholesterol. Both the proper expression of ABCA1 in the plasma membrane and the internalization along with apoA-I are required for function.... Ausführliche Beschreibung

1. Person: Sarkadi Balázs
Weitere Personen: Váradi András; Andrikovics Hajnalka; Német Katalin; Szabó Katalin; Hegyi Zoltán; Kasza Ildikó; Homolya László
Quelle: In BMC Cell Biology (01.12.2009)
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Format: Online-Artikel
Sprache: English
Veröffentlicht: 2009
Beschreibung: Online-Ressource
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  Creative Commons License Source: Directory of Open Access Journals (DOAJ).
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024 7 |a 10.1186/1471-2121-10-93  |2 doi 
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041 0 |a eng 
100 0 |a Sarkadi Balázs 
245 1 0 |a Model system for the analysis of cell surface expression of human ABCA1  |h Elektronische Ressource 
300 |a Online-Ressource 
520 |a Abstract Background The ABCA1 protein plays a pivotal role in reverse cholesterol transport, by mediating the generation of HDL particles and removing cellular cholesterol. Both the proper expression of ABCA1 in the plasma membrane and the internalization along with apoA-I are required for function. Therefore, we developed a model system to investigate the effect of clinically relevant drugs on the cell surface appearance of ABCA1. Results By retroviral transduction system, we established stable mammalian cell lines expressing functional and non-functional ABCA1 variants, tagged with an extracellular hemagglutinin epitope. After characterization of the expression, proper localization and function of different ABCA1 variants, we followed quantitatively their cell surface expression by immunofluorescent staining, using flow cytometry. As expected, we found increased cell surface expression of ABCA1 after treatment with a calpain inhibitor, and observed a strong decrease in plasma membrane ABCA1 expression upon treatment with a trans-Golgi transport inhibitor, Brefeldin A. We tested cholesterol level lowering drugs and other potential inhibitors of ABCA1. Here we demonstrate that ezetimibe affects ABCA1 cell surface expression only in the case of a functional ABCA1. Conclusions Our model system allows a quantitative detection of cell surface expression of ABCA1, screening of substrates or specific inhibitors, and investigating transport regulation. 
700 0 |a Váradi András 
700 0 |a Andrikovics Hajnalka 
700 0 |a Német Katalin 
700 0 |a Szabó Katalin 
700 0 |a Hegyi Zoltán 
700 0 |a Kasza Ildikó 
700 0 |a Homolya László 
773 0 8 |i In  |t BMC Cell Biology  |g  (01.12.2009)  |w (DE-601)DOAJ000053910  |x 1471-2121 
856 4 0 |u http://dx.doi.org/10.1186/1471-2121-10-93 
856 4 0 |y DOAJ  |u https://doaj.org/article/a1928f46c7d74442bbed595a76cc415e 
856 4 0 |u http://www.biomedcentral.com/1471-2121/10/93 
856 4 0 |u https://doaj.org/toc/1471-2121 
912 |a GBV_DOAJ 
951 |a AR 
952 |j 2009  |b 01  |c 12 

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