title: Subnanometre-resolution electron cryomicroscopy structure of a heterodimeric ABC exporter.
abstract: ATP-binding cassette (ABC) transporters translocate substrates across cell membranes, using energy harnessed from ATP binding and hydrolysis at their nucleotide-binding domains. ABC exporters are present both in prokaryotes and eukaryotes, with examples implicated in multidrug resistance of pathogens and cancer cells, as well as in many human diseases. TmrAB is a heterodimeric ABC exporter from the thermophilic Gram-negative eubacterium Thermus thermophilus; it is homologous to various multidrug transporters and contains one degenerate site with a non-catalytic residue next to the Walker B motif. Here we report a subnanometre-resolution structure of detergent-solubilized TmrAB in a nucleotide-free, inward-facing conformation by single-particle electron cryomicroscopy. The reconstructions clearly resolve characteristic features of ABC transporters, including helices in the transmembrane domain and nucleotide-binding domains. A cavity in the transmembrane domain is accessible laterally from the cytoplasmic side of the membrane as well as from the cytoplasm, indicating that the transporter lies in an inward-facing open conformation. The two nucleotide-binding domains remain in contact via their carboxy-terminal helices. Furthermore, comparison between our structure and the crystal structures of other ABC transporters suggests a possible trajectory of conformational changes that involves a sliding and rotating motion between the two nucleotide-binding domains during the transition from the inward-facing to outward-facing conformations.
authors.affiliation: Department of Pharmaceutical Chemistry, University of California San Francisco, 600 16th Street, San Francisco, California 94158, USA.
authors.forename: JungMin
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, California 94158, USA.
authors.forename: Shenping
authors.initials: S
authors.lastname: Wu
authors.affiliation: Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, California 94158, USA.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Tomasiak
authors.affiliation: Institute of Biochemistry, Biocenter, Goethe-University Frankfurt, Max-von-Laue-Strasse 9, D-60438 Frankfurt am Main, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Mergel
authors.affiliation: Department of Pharmaceutical Chemistry, University of California San Francisco, 600 16th Street, San Francisco, California 94158, USA.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Winter
authors.affiliation: Institute of Biochemistry, Biocenter, Goethe-University Frankfurt, Max-von-Laue-Strasse 9, D-60438 Frankfurt am Main, Germany.
authors.forename: Sebastian B
authors.initials: SB
authors.lastname: Stiller
authors.affiliation: Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, California 94158, USA.
authors.forename: Yaneth
authors.initials: Y
authors.lastname: Robles-Colmanares
authors.affiliation: 1] Department of Pharmaceutical Chemistry, University of California San Francisco, 600 16th Street, San Francisco, California 94158, USA [2] Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, California 94158, USA.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Stroud
authors.affiliation: 1] Institute of Biochemistry, Biocenter, Goethe-University Frankfurt, Max-von-Laue-Strasse 9, D-60438 Frankfurt am Main, Germany [2] Cluster of Excellence - Macromolecular Complexes, Goethe-University Frankfurt, Max-von-Laue-Strasse 9, D-60438 Frankfurt am Main, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Tampé
authors.affiliation: Department of Pharmaceutical Chemistry, University of California San Francisco, 600 16th Street, San Francisco, California 94158, USA.
authors.forename: Charles S
authors.initials: CS
authors.lastname: Craik
authors.affiliation: Department of Biochemistry and Biophysics, University of California San Francisco, 600 16th Street, San Francisco, California 94158, USA.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Cheng
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000276:immunology / Q000648:ultrastructure*; D000941:Antigens / Q000737:chemistry / Q000276:immunology; D001665:Binding Sites; D020285:Cryoelectron Microscopy; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D009711:Nucleotides / Q000378:metabolism; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D012399:Rotation; D016963:Thermus thermophilus / Q000737:chemistry*
chemical_list: D018528:ATP-Binding Cassette Transporters; D000941:Antigens; D009711:Nucleotides