Modulation of LAT1 (SLC7A5) transporter activity and stability by membrane cholesterol.



Dickens, D ORCID: 0000-0001-8295-0752, Chiduza, GN ORCID: 0000-0003-1037-5749, Wright, GSA ORCID: 0000-0002-3756-9634, Pirmohamed, M ORCID: 0000-0002-7534-7266, Antonyuk, SV ORCID: 0000-0002-2779-9946 and Hasnain, SS
(2017) Modulation of LAT1 (SLC7A5) transporter activity and stability by membrane cholesterol. Scientific Reports, 7 (1). 43580-.

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Abstract

LAT1 (SLC7A5) is a transporter for both the uptake of large neutral amino acids and a number of pharmaceutical drugs. It is expressed in numerous cell types including T-cells, cancer cells and brain endothelial cells. However, mechanistic knowledge of how it functions and its interactions with lipids are unknown or limited due to inability of obtaining stable purified protein in sufficient quantities. Our data show that depleting cellular cholesterol reduced the Vmax but not the Km of the LAT1 mediated uptake of a model substrate into cells (L-DOPA). A soluble cholesterol analogue was required for the stable purification of the LAT1 with its chaperon CD98 (4F2hc,SLC3A2) and that this stabilised complex retained the ability to interact with a substrate. We propose cholesterol interacts with the conserved regions in the LAT1 transporter that have been shown to bind to cholesterol/CHS in Drosophila melanogaster dopamine transporter. In conclusion, LAT1 is modulated by cholesterol impacting on its stability and transporter activity. This novel finding has implications for other SLC7 family members and additional eukaryotic transporters that contain the LeuT fold.

Item Type: Article
Uncontrolled Keywords: Humans, Levodopa, Cholesterol, Membrane Lipids, Amino Acids, Large Neutral Amino Acid-Transporter 1, Gene Expression, Binding Sites, Amino Acid Sequence, Amino Acid Motifs, Protein Conformation, Protein Binding, Biological Transport, Kinetics, Models, Molecular, Lipid Metabolism, Protein Interaction Domains and Motifs, Fusion Regulatory Protein-1
Depositing User: Symplectic Admin
Date Deposited: 10 Mar 2017 11:50
Last Modified: 19 Jan 2023 07:14
DOI: 10.1038/srep43580
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3006321