Universität zu Köln

Gründemann, Dirk ORCID: 0000-0003-0914-0299, Flögel, Svenja, Tust, Maurice, Boussettaoui, Samira and Fischer, Dietmar (2025). Enhancing transporter activity in heterologous expression systems with SAHA : a 2500‐times more potent and odorless alternative to butyrate. FEBS Open Bio, 15 (6). pp. 906-913. Wiley. ISSN 2211-5463

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Abstract

The functional characterization of plasma membrane transport proteins often relies on their heterologous expression in cultured cells. However, some transporters exhibit low activity, hindering meaningful functional assays. Heterologous expression is usually based on strong viral promoters which in living cells are prone to promoter silencing, a major problem. Here, we investigated the efficacy of low‐cost histone deacetylase (HDAC) inhibitors in enhancing transporter activity, comparing the established sodium butyrate (the sodium salt of butyric acid) with valproate/valproic acid (VPA) and suberoylanilide hydroxamic acid (SAHA, also known as vorinostat). Using 293 cells stably transfected with pEBTet plasmids containing the CMV promotor to express the transporters SLC16A9, SLC22A15, and OATP1A2, we measured substrate efflux or uptake via LC–MS/MS following overnight preincubation with the HDAC inhibitors. All three compounds markedly stimulated transporter activity. VPA was less effective than butyrate but still surpassed control conditions. SAHA was cytotoxic at 6 μ m , but at 2 μ m , the enhancement was consistently comparable to 5 m m butyrate. Additionally, SAHA was more cost‐effective and devoid of the repulsive odor characteristic of butyrate. Our findings advocate for replacing butyrate with SAHA to enhance heterologously expressed transporter activity. This offers a more efficient and user‐friendly alternative for functional assays.

Item Type:	Article
Creators:	Creators Email ORCID ORCID Put Code Gründemann, Dirk UNSPECIFIED https://orcid.org/0000-0003-0914-0299 UNSPECIFIED Flögel, Svenja UNSPECIFIED UNSPECIFIED UNSPECIFIED Tust, Maurice UNSPECIFIED UNSPECIFIED UNSPECIFIED Boussettaoui, Samira UNSPECIFIED UNSPECIFIED UNSPECIFIED Fischer, Dietmar UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-791849
Identification Number:	10.1002/2211-5463.70015
Journal or Publication Title:	FEBS Open Bio
Volume:	15
Number:	6
Page Range:	pp. 906-913
Number of Pages:	8
Date:	7 March 2025
Publisher:	Wiley
ISSN:	2211-5463
Language:	English
Faculty:	Faculty of Medicine
Divisions:	Faculty of Medicine > Pharmakologie > Instítut für Pharmakologie
Subjects:	Medical sciences Medicine
['eprint_fieldname_oa_funders' not defined]:	Publikationsfonds UzK
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/79184