A5040243630- Cellular Mechanics and Interactions
- Cell Adhesion Molecules Research
- Molecular spectroscopy and chirality
- RNA Research and Splicing
- NMR spectroscopy and applications
- Advanced NMR Techniques and Applications
- Cellular transport and secretion
- Microtubule and mitosis dynamics
- S100 Proteins and Annexins
- Protein Kinase Regulation and GTPase Signaling
- Enzyme Structure and Function
- Ubiquitin and proteasome pathways
- Photoreceptor and optogenetics research
- Protein Structure and Dynamics
- Metabolomics and Mass Spectrometry Studies
- Caveolin-1 and cellular processes
- Nuclear Structure and Function
- Biochemical and Structural Characterization
- Mitochondrial Function and Pathology
- Glycosylation and Glycoproteins Research
- Cardiomyopathy and Myosin Studies
- Computational Drug Discovery Methods
- Force Microscopy Techniques and Applications
- Proteoglycans and glycosaminoglycans research
- Protease and Inhibitor Mechanisms
University of Liverpool
2016-2025
University of Leicester
1999-2009
University of Nottingham
2006
University of Liège
1999
Institute of Bioorganic Chemistry
1984-1992
Russian Academy of Sciences
1992
The structure of [Val1]gramicidin A incorporated into sodium dodecyl-d25 sulphate micelles has been studied by two-dimensional proton NMR spectroscopy. Analysis nuclear Overhauser effects, spin-spin couplings and solvent accessibility NH groups show that the conformation Na+ complex gramicidin in detergent micelles, which many ways mimic phospholipid bilayer biomembranes, is an N-terminal to (head-to-head) dimer (Formula: see text) formed two right-handed, single-stranded beta 6.3 helices...
Talin activates integrins, couples them to F-actin, and recruits vinculin focal adhesions (FAs). Here, we report the structural characterization of talin rod: 13 helical bundles (R1–R13) organized into a compact cluster four-helix (R2–R4) within linear chain five-helix bundles. Nine contain vinculin-binding sites (VBS); R2R3 are atypical, with each containing two VBS. also binds synergistically RIAM, Rap1 effector involved in integrin activation. Biochemical data show that RIAM binding is...
Abstract The link between extracellular-matrix-bound integrins and intracellular F-actin is essential for cell spreading migration. Here, we demonstrate how the actin-binding proteins talin vinculin cooperate to provide this link. By expressing structure-based mutants in null cells, show that while C-terminal site (ABS3) required adhesion complex assembly, central ABS2 focal (FA) maturation. Thus, although support spreading, cells lack FAs, fail polarize exert reduced force on surrounding...
The interaction between the cytoskeletal proteins talin and vinculin plays a key role in integrin-mediated cell adhesion migration. Three binding sites (VBS1-3) have previously been identified rod using yeast two-hybrid assay. To extend these studies, we spot-synthesized series of peptides spanning all alpha-helical regions predicted for eight additional VBSs, two which overlap functional rod, including integrin site C-terminal actin site. VBS alpha-helices bind to hydrophobic cleft...
SummaryFERM domains are found in a diverse superfamily of signaling and adaptor proteins at membrane interfaces. They typically consist three separately folded (F1, F2, F3) compact cloverleaf structure. The crystal structure the N-terminal head integrin-associated cytoskeletal protein talin reported here reveals novel FERM domain with linear arrangement, plus an additional F0 packed against F1. While F3 binds β-integrin tails, basic residues F1 F2 required for association integrin...
NADPH-cytochrome P450 reductase (CPR), a diflavin reductase, plays key role in the mammalian mono-oxygenase system. In its crystal structure, two flavins are close together, positioned for interflavin electron transfer but not to cytochrome P450. A number of lines evidence suggest that domain motion is important action enzyme. We report NMR and small-angle x-ray scattering experiments addressing directly question organization human CPR. Comparison (1)H-(15)N heteronuclear single quantum...
Talin is a large flexible rod-shaped protein that activates the integrin family of cell adhesion molecules and couples them to cytoskeletal actin. It exists in both globular extended conformations, an intramolecular interaction between N-terminal F3 FERM subdomain C-terminal part talin rod contributes autoinhibited form molecule. Here, we report solution structure primary binding domain within region use intermolecular nuclear Overhauser effects determine complex. The (residues 1655–1822)...
Cell migration requires coordination between integrin-mediated cell adhesion to the extracellular matrix and force applied sites. Talin plays a key role in coupling integrin receptors actomyosin contractile machinery, while deleted liver cancer 1 (DLC1) is Rho GAP that binds talin regulates Rho, therefore contractility. We show LD motif of DLC1 forms helix four-helix bundle R8 domain canonical triple-helix arrangement. demonstrate same surface interacts with paxillin LD1 LD2 motifs. identify...
The adaptor protein talin serves both to activate the integrin family of cell adhesion molecules and couple integrins actin cytoskeleton. Integrin activation has been shown involve binding FERM domain membrane proximal sequences in cytoplasmic β-subunit. However, a second integrin-binding site (IBS2) identified near C-terminal end rod. Here we report crystal structure IBS2 (residues 1974-2293), which comprises two five-helix bundles, "IBS2-A" (1974-2139) "IBS2-B" (2140-2293), connected by...
The integrin family of heterodimeric cell adhesion molecules exists in both low- and high-affinity states, activation requires binding the talin FERM (four-point-one, ezrin, radixin, moesin) domain to membrane-proximal sequences β-integrin cytoplasmic domain. However, it has recently become apparent that kindlin proteins is also essential for talin-induced activation. domains are typically composed F1, F2, F3 domains, but atypical contains a large insert F1 preceded by previously...
Talin is an adaptor protein that couples integrins to F-actin. Structural studies show the N-terminal talin head contains atypical FERM domain, whereas N- and C-terminal parts of rod include a series α-helical bundles. However, determining structure central part has proved problematic. Residues 1359–1659 are homologous MESDc1 gene product, we therefore expressed this region in Escherichia coli. The crystal shows unique fold comprised 5- 4-helix bundle. 5-helix bundle composed nonsequential...
Talin, vinculin, and paxillin are core components of the dynamic link between integrins actomyosin. Here, we study mechanisms that mediate their activation association using a mitochondrial-targeting assay, structure-based mutants, advanced microscopy. As expected, full-length vinculin talin autoinhibited do not interact with each other. However, contrary to previous models propose critical role for forces driving talin-vinculin association, our data show force-independent relief...
Talin is an essential component of focal adhesions that couples β-integrin cytodomains to F-actin and provides a scaffold for signaling proteins. Recently, the integrin β3 cytodomain phosphatidylinositol phosphate (PIP) kinase type 1γ (a 4,5-bisphosphate-synthesizing enzyme) were shown bind talin FERM domain (subdomain F3). We have characterized PIP kinase-binding site by NMR using 15N-labeled F2F3 polypeptide. A peptide containing minimal talin-binding formed 1:1 complex with F2F3, causing...