Projection structure of a member of the amino acid/polyamine/organocation transporter superfamily

Casagrande, Fabio; Ratera, Merce; Schenk, Andreas D; Chami, Mohamed; Valencia, Eva; Lopez, Jesus Maria; Torrents, David; Engel, Andreas; Palacin, Manuel; Fotiadis, Dimitrios (2008). Projection structure of a member of the amino acid/polyamine/organocation transporter superfamily. Journal of biological chemistry, 283(48), pp. 33240-8. Bethesda, Md.: American Society for Biochemistry and Molecular Biology 10.1074/jbc.M806917200

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The L-arginine/agmatine antiporter AdiC is a key component of the arginine-dependent extreme acid resistance system of Escherichia coli. Phylogenetic analysis indicated that AdiC belongs to the amino acid/polyamine/organocation (APC) transporter superfamily having sequence identities of 15-17% to eukaryotic and human APC transporters. For functional and structural characterization, we cloned, overexpressed, and purified wild-type AdiC and the point mutant AdiC-W293L, which is unable to bind and consequently transport L-arginine. Purified detergent-solubilized AdiC particles were dimeric. Reconstitution experiments yielded two-dimensional crystals of AdiC-W293L diffracting beyond 6 angstroms resolution from which we determined the projection structure at 6.5 angstroms resolution. The projection map showed 10-12 density peaks per monomer and suggested mainly tilted helices with the exception of one distinct perpendicular membrane spanning alpha-helix. Comparison of AdiC-W293L with the projection map of the oxalate/formate antiporter from Oxalobacter formigenes, a member from the major facilitator superfamily, indicated different structures. Thus, two-dimensional crystals of AdiC-W293L yielded the first detailed view of a transport protein from the APC superfamily at sub-nanometer resolution.

Item Type:

Journal Article (Original Article)


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

UniBE Contributor:

Fotiadis, Dimitrios José






American Society for Biochemistry and Molecular Biology




Factscience Import

Date Deposited:

04 Oct 2013 15:03

Last Modified:

04 May 2014 23:19

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URI: (FactScience: 106006)

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