Impact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter

The Proton-Coupled Folate Transporter (PCFT) is a transmembrane transport protein that controls the absorption of dietary folates in the small intestine. PCFT also mediates uptake of chemotherapeutically used antifolates into tumor cells. PCFT has been identified within lipid rafts observed in phosp...

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Autores principales:	Hoa Quynh Do, Carla M. Bassil, Elizabeth I. Andersen, Michaela Jansen
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/dfc66f783ce5431589ec3cad178398fd
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spelling	oai:doaj.org-article:dfc66f783ce5431589ec3cad178398fd2021-11-25T06:19:36ZImpact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter1932-6203https://doaj.org/article/dfc66f783ce5431589ec3cad178398fd2021-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8601550/?tool=EBIhttps://doaj.org/toc/1932-6203The Proton-Coupled Folate Transporter (PCFT) is a transmembrane transport protein that controls the absorption of dietary folates in the small intestine. PCFT also mediates uptake of chemotherapeutically used antifolates into tumor cells. PCFT has been identified within lipid rafts observed in phospholipid bilayers of plasma membranes, a micro environment that is altered in tumor cells. The present study aimed at investigating the impact of different lipids within Lipid-protein nanodiscs (LPNs), discoidal lipid structures stabilized by membrane scaffold proteins, to yield soluble PCFT expression in an E. coli lysate-based cell-free transcription/translation system. In the absence of detergents or lipids, we observed PCFT quantitatively as precipitate in this system. We then explored the ability of LPNs to support solubilized PCFT expression when present during in-vitro translation. LPNs consisted of either dimyristoyl phosphatidylcholine (DMPC), palmitoyl-oleoyl phosphatidylcholine (POPC), or dimyristoyl phosphatidylglycerol (DMPG). While POPC did not lead to soluble PCFT expression, both DMPG and DMPC supported PCFT translation directly into LPNs, the latter in a concentration dependent manner. The results obtained through this study provide insights into the lipid preferences of PCFT. Membrane-embedded or solubilized PCFT will enable further studies with diverse biophysical approaches to enhance the understanding of the structure and molecular mechanism of folate transport through PCFT.Hoa Quynh DoCarla M. BassilElizabeth I. AndersenMichaela JansenPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Hoa Quynh Do Carla M. Bassil Elizabeth I. Andersen Michaela Jansen Impact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter
description	The Proton-Coupled Folate Transporter (PCFT) is a transmembrane transport protein that controls the absorption of dietary folates in the small intestine. PCFT also mediates uptake of chemotherapeutically used antifolates into tumor cells. PCFT has been identified within lipid rafts observed in phospholipid bilayers of plasma membranes, a micro environment that is altered in tumor cells. The present study aimed at investigating the impact of different lipids within Lipid-protein nanodiscs (LPNs), discoidal lipid structures stabilized by membrane scaffold proteins, to yield soluble PCFT expression in an E. coli lysate-based cell-free transcription/translation system. In the absence of detergents or lipids, we observed PCFT quantitatively as precipitate in this system. We then explored the ability of LPNs to support solubilized PCFT expression when present during in-vitro translation. LPNs consisted of either dimyristoyl phosphatidylcholine (DMPC), palmitoyl-oleoyl phosphatidylcholine (POPC), or dimyristoyl phosphatidylglycerol (DMPG). While POPC did not lead to soluble PCFT expression, both DMPG and DMPC supported PCFT translation directly into LPNs, the latter in a concentration dependent manner. The results obtained through this study provide insights into the lipid preferences of PCFT. Membrane-embedded or solubilized PCFT will enable further studies with diverse biophysical approaches to enhance the understanding of the structure and molecular mechanism of folate transport through PCFT.
format	article
author	Hoa Quynh Do Carla M. Bassil Elizabeth I. Andersen Michaela Jansen
author_facet	Hoa Quynh Do Carla M. Bassil Elizabeth I. Andersen Michaela Jansen
author_sort	Hoa Quynh Do
title	Impact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter
title_short	Impact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter
title_full	Impact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter
title_fullStr	Impact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter
title_full_unstemmed	Impact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter
title_sort	impact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/dfc66f783ce5431589ec3cad178398fd
work_keys_str_mv	AT hoaquynhdo impactofnanodisclipidcompositiononcellfreeexpressionofprotoncoupledfolatetransporter AT carlambassil impactofnanodisclipidcompositiononcellfreeexpressionofprotoncoupledfolatetransporter AT elizabethiandersen impactofnanodisclipidcompositiononcellfreeexpressionofprotoncoupledfolatetransporter AT michaelajansen impactofnanodisclipidcompositiononcellfreeexpressionofprotoncoupledfolatetransporter
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Impact of nanodisc lipid composition on cell-free expression of proton-coupled folate transporter

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