title: The structure of the C-terminal actin-binding domain of talin.
abstract: Talin is a large dimeric protein that couples integrins to cytoskeletal actin. Here, we report the structure of the C-terminal actin-binding domain of talin, the core of which is a five-helix bundle linked to a C-terminal helix responsible for dimerisation. The NMR structure of the bundle reveals a conserved surface-exposed hydrophobic patch surrounded by positively charged groups. We have mapped the actin-binding site to this surface and shown that helix 1 on the opposite side of the bundle negatively regulates actin binding. The crystal structure of the dimerisation helix reveals an antiparallel coiled-coil with conserved residues clustered on the solvent-exposed face. Mutagenesis shows that dimerisation is essential for filamentous actin (F-actin) binding and indicates that the dimerisation helix itself contributes to binding. We have used these structures together with small angle X-ray scattering to derive a model of the entire domain. Electron microscopy provides direct evidence for binding of the dimer to F-actin and indicates that it binds to three monomers along the long-pitch helix of the actin filament.
authors.affiliation: Department of Biochemistry, University of Leicester, Leicester, UK.
authors.forename: Alexandre R
authors.initials: AR
authors.lastname: Gingras
authors.forename: Neil
authors.initials: N
authors.lastname: Bate
authors.forename: Benjamin T
authors.initials: BT
authors.lastname: Goult
authors.forename: Larnele
authors.initials: L
authors.lastname: Hazelwood
authors.forename: Ilona
authors.initials: I
authors.lastname: Canestrelli
authors.forename: J Günter
authors.initials: JG
authors.lastname: Grossmann
authors.forename: HongJun
authors.initials: H
authors.lastname: Liu
authors.forename: Nicholas S M
authors.initials: NS
authors.lastname: Putz
authors.forename: Gordon C K
authors.initials: GC
authors.lastname: Roberts
authors.forename: Niels
authors.initials: N
authors.lastname: Volkmann
authors.forename: Dorit
authors.initials: D
authors.lastname: Hanein
authors.forename: Igor L
authors.initials: IL
authors.lastname: Barsukov
authors.forename: David R
authors.initials: DR
authors.lastname: Critchley
mesh_terms: D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D018360:Crystallography, X-Ray; D019281:Dimerization; D007091:Image Processing, Computer-Assisted; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D016608:Talin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D016608:Talin