2D and 3D crystallization of a bacterial homologue of human vitamin C membrane transport proteins

Jeckelmann, Jean-Marc; Harder, Daniel; Ucurum Fotiadis, Zöhre; Fotiadis, Dimitrios José (2014). 2D and 3D crystallization of a bacterial homologue of human vitamin C membrane transport proteins. Journal of structural biology, 188(1), pp. 87-91. Elsevier 10.1016/j.jsb.2014.08.004

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Most organisms are able to synthesize vitamin C whereas humans are not. In order to contribute to the elucidation of the molecular working mechanism of vitamin C transport through biological membranes, we cloned, overexpressed, purified, functionally characterized, and 2D- and 3D-crystallized a bacterial protein (UraDp) with 29% of amino acid sequence identity to the human sodium-dependent vitamin C transporter 1 (SVCT1). Ligand-binding experiments by scintillation proximity assay revealed that uracil is a substrate preferably bound to UraDp. For structural analysis, we report on the production of tubular 2D crystals and present a first projection structure of UraDp from negatively stained tubes. On the other hand the successful growth of UraDp 3D crystals and their crystallographic analysis is described. These 3D crystals, which diffract X-rays to 4.2Å resolution, pave the way towards the high-resolution crystal structure of a bacterial homologue with high amino acid sequence identity to human SVCT1.

Item Type:

Journal Article (Original Article)

Division/Institute:

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

UniBE Contributor:

Jeckelmann, Jean-Marc; Harder, Daniel; Ucurum Fotiadis, Zöhre and Fotiadis, Dimitrios José

Subjects:

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

ISSN:

1047-8477

Publisher:

Elsevier

Language:

English

Submitter:

Barbara Järmann-Bangerter

Date Deposited:

02 Apr 2015 08:35

Last Modified:

02 Sep 2015 10:21

Publisher DOI:

10.1016/j.jsb.2014.08.004

PubMed ID:

25160726

Uncontrolled Keywords:

Membrane protein, Projection structure, Scintillation proximity assay, Three-dimensional crystal, Transmission electron microscopy, Transport protein, Two-dimensional crystal, X-ray crystallography

BORIS DOI:

10.7892/boris.65711

URI:

https://boris.unibe.ch/id/eprint/65711

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