3DYJ

Crystal Structure of the R11R12 Domains of Talin

Summary for 3DYJ

• Entry DOI: 10.2210/pdb3dyj/pdb
• Descriptor: TALIN-1 (2 entities in total)
• Functional Keywords: talin, helix bundles, cytoskeletal protein, integrin-bindin site, ibs2, cell membrane, cell projection, cytoplasm, cytoskeleton, membrane, phosphoprotein, structural protein
• Biological source: Mus musculus (mouse)
• Cellular location: Cell projection, ruffle membrane; Peripheral membrane protein; Cytoplasmic side (By similarity): P26039
• Total number of polymer chains: 2
• Total formula weight: 70464.41

Authors

Gingras, A.R.,Joyce, M.G.,Critchley, D.R.,Emsley, J. (deposition date: 2008-07-28, release date: 2009-01-27, Last modification date: 2024-10-09)

Primary citation

Gingras, A.R.,Ziegler, W.H.,Bobkov, A.A.,Joyce, M.G.,Fasci, D.,Himmel, M.,Rothemund, S.,Ritter, A.,Grossmann, J.G.,Patel, B.,Bate, N.,Goult, B.T.,Emsley, J.,Barsukov, I.L.,Roberts, G.C.,Liddington, R.C.,Ginsberg, M.H.,Critchley, D.R.
Structural determinants of integrin binding to the talin rod
J.Biol.Chem., 284:8866-8876, 2009

PubMed Abstract: The adaptor protein talin serves both to activate the integrin family of cell adhesion molecules and to couple integrins to the actin cytoskeleton. Integrin activation has been shown to involve binding of the talin FERM domain to membrane proximal sequences in the cytoplasmic domain of the integrin beta-subunit. However, a second integrin-binding site (IBS2) has been identified near the C-terminal end of the talin rod. Here we report the crystal structure of IBS2 (residues 1974-2293), which comprises two five-helix bundles, "IBS2-A" (1974-2139) and "IBS2-B" (2140-2293), connected by a continuous helix with a distinct kink at its center that is stabilized by side-chain H-bonding. Solution studies using small angle x-ray scattering and NMR point to a fairly flexible quaternary organization. Using pull-down and enzyme-linked immunosorbent assays, we demonstrate that integrin binding requires both IBS2 domains, as does binding to acidic phospholipids and robust targeting to focal adhesions. We have defined the membrane proximal region of the integrin cytoplasmic domain as the major binding region, although more membrane distal regions are also required for strong binding. Alanine-scanning mutagenesis points to an important electrostatic component to binding. Thermal unfolding experiments show that integrin binding induces conformational changes in the IBS2 module, which we speculate are linked to vinculin and membrane binding.

PubMed: 19176533
DOI: 10.1074/jbc.M805937200

Experimental method

X-RAY DIFFRACTION (1.85 Å)