Expression, purification and low-resolution structure of human vitamin C transporter SVCT1 (SLC23A1)

Boggavarapu, Rajendra; Jeckelmann, Jean-Marc; Harder, Daniel; Schneider, Philipp; Ucurum, Zöhre; Hediger, Matthias; Fotiadis, Dimitrios (2013). Expression, purification and low-resolution structure of human vitamin C transporter SVCT1 (SLC23A1). PLoS ONE, 8(10), e76427. Public Library of Science 10.1371/journal.pone.0076427

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Expression and purification of human membrane proteins for structural studies represent a great challenge. This is because micro- to milligram amounts of pure isolated protein are required. To this aim, we successfully expressed the human vitamin C transporter-1 (hSVCT1; SLC23A1) in Xenopus laevis oocytes and isolated highly pure protein in microgram amounts. Recombinant hSVCT1 was functional when expressed in oocytes and glycosylated. Structural analysis of purified hSVCT1 by transmission electron microscopy and single particle analysis unveiled its shape, dimensions and low-resolution structure as well as the existence of a major monomeric and minor dimeric population. Chemical crosslinking of isolated oocyte membranes containing expressed hSVCT1 indicated similar oligomeric states of hSVCT1 in lipid bilayers. This work reports the first purification and structural analysis of a human SVCT protein and opens the way for future functional and structural studies using purified hSVCT1.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Biochemistry and Molecular Medicine

UniBE Contributor:

Boggavarapu, Venkata Ravi Naga Rajendr; Jeckelmann, Jean-Marc; Harder, Daniel; Schneider, Philipp; Ucurum Fotiadis, Zöhre; Hediger, Matthias and Fotiadis, Dimitrios José

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

1932-6203

Publisher:

Public Library of Science

Language:

English

Submitter:

Patrizia Catucci

Date Deposited:

01 May 2014 12:42

Last Modified:

05 Dec 2014 04:35

Publisher DOI:

10.1371/journal.pone.0076427

PubMed ID:

24124560

BORIS DOI:

10.7892/boris.43402

URI:

https://boris.unibe.ch/id/eprint/43402

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