| Talin (C-terminal-domain and actin |
EM, SAXS, docking and helical reconstruction |
Completed - Gingras et al., 2008 |
Hanein (Liddington, Critchley, Volkmann) |
| Alpha-actinin beta1-integrin complex |
2D crystallization on lipid monolayers |
Completed - Kelly & Taylor, 2005 |
Taylor |
| Alpha-actinin2 (chicken smooth muscle) |
2D crystallization on lipid monolayers |
Completed - Liu, et al, 2004 |
Taylor (J. Liu, D. Taylor) |
| Arp2/3 branch structure |
Cryo EM |
Completed - 2D structure completed Volkmann et al., 2001 |
Hanein (Volkmann, Li, Pollard) |
| Metavinculin C-terminal domain |
X-Ray crystallography, NMR |
Completed - Bakolitsa et al., 1999
|
Liddington |
| Lamellipodia leading edge (Ena/VASP proteins) |
Correlative light microscopy-electron microscopy |
Completed - Bear et al., 2002 |
Borisy (Gertler) |
| WASP and Scar - Drosophila actin nucleation |
Correlative light microscopy-electron microscopy |
Completed - Biyasheva et al., 2004 |
Borisy (Baum) |
| Actin comet tail |
Correlative light microscopy-electron microscopy |
Completed - Cameron et al., 2001 |
Borisy (Theriot) |
| PIPKIg-talin(196-400) |
X-Ray crystallography, NMR |
Completed - de Pereda et al., 2004 |
Liddington (Campbell) |
Tropomyosin  and the leading edge |
conventional EM |
Completed - Gupton et al. 2005
|
Hanein (Waterman-Storer) |
| Vinculin-tail actin |
EM, tomography and docking |
Completed - Janssen et al., 2006 |
Hanein (Volkmann, Liddington) |
| Rickettsia - structure of tail mediated by WASP-like protein |
Correlative light microscopy-electron microscopy |
Completed - Jeng et al., 2004 |
Borisy (Welch) |
| Alpha-actinin beta1-integrin-vinculin1-258 complex |
2D crystallization on lipid monolayers |
Completed - Kelly, et al., 2006 |
Taylor (Liddington) |
| Talin (Ferm-domain) and actin |
EM and helical reconstruction |
Completed - Lee et al, 2004 |
Hanein (Liddington, Critchley, Volkmann) |
| Arp2/3 activation (mutants) |
EM, single particle reconstruction and docking |
Completed - Martin et al., 2005 |
Hanein (Drubin, Volkmann) |
| Tensin SH2 and PTB domains |
X-Ray crystallography, NMR |
Completed - McCleverty et al. (2007) |
Liddington |
| HIV-cytoskeletal association (microtubules) |
Correlative light microscopy-electron microscopy |
Completed - McDonald et al., 2002 |
Borisy (Hope) |
| Alpha-actinin cross-links |
2D crystallization on lipid monolayers |
Completed - Taylor et al 2007 |
Taylor |
| Actin filaments orientational order in vivo |
Correlative light microscopy-electron microscopy |
Completed - Verkhovsky et al., 2003 |
Borisy (Meister) |
| Myosin and actin |
Cryo EM, helical reconstruction, docking |
Completed - Volkmann et al 2000, Volkmann et al 2003, Volkmann & Hanein 2000.Volkmann, et al., 2007,Trybus et al., 2007,Lowey et al., 2007 |
Hanein (Volkmann, Lowey, Trybus, DeRosier) |
| Actin fimbrin crosslink |
EM and bioinformatics |
Completed - completed Volkmann et al., 2001 |
Hanein (Volkmann) |
| Talin head domain:beta 3 integrin tail complex |
X-Ray crystallography, NMR |
Completed - crystallized and solved Garcia-Alvarez et al., 2003 |
Liddington (Campbell) |
| FAT domain of Focal Adhesion Kinase |
X-Ray crystallography, NMR |
Completed - online and Hayashi et al., 2002 |
Liddington |
| Vinculin (full length) |
X-Ray crystallography |
Completed - Structure determined Bakolitsa et al., 2004 (Bakolitsa et al., 1999 Saunders et al., 2006 and Ziegler et al, 2006.. |
Liddington |
| Alpha-actinin2/F-actin rafts |
lipid monolayer, negative stain, single particle image processing |
Completed - in press - J. Mol. Biol. |
Taylor |
| Mapping of molecular sub-domains |
Multi color labeling; Clustering analysis |
Completed, publication in progress |
Geiger |
| Osteoclast structure |
Multi color labeling |
Completed, publication in progress |
Geiger |
| Cell-matrix interactions |
Multi color labeling |
Completed, publication in progress |
Geiger (Hanein) |
| Leading edge |
VMP, Cryo EM and conventional EM |
Completed, publication in progress |
Hanein |
| Fibronectin fibrillogenises |
EM, light and EM labeling, cell culture |
Completed, publication in progress |
Hanein (Geiger, Engel, Ruoslahti) |
| F-actin-villin (with David DeRosier) |
Electron Tomography |
Completed, publication in progress |
Taylor |
| alphaIIb-beta3 integrin structure in the membrane |
Electron Tomography |
Completed, publication in progress |
Taylor |
| Focal adhesion structure |
Correlative light microscopy-electron microscopy |
In progress |
Geiger (Hanein) |
| Organization of zyxin and alpha-actinin in focal adhesions and stress fibers |
Correlative light microscopy-electron microscopy |
In progress |
Geiger (Hanein, Taylor) |
| Podosome structure |
VMP and Cryo EM |
In progress |
Hanein (Geiger) |
| Focal adhesion structure in-situ |
VMP, light and EM, labeling, Cryo EM and tomography |
In progress |
Hanein (Geiger) |
| Vasp-actin |
EM, tomography, single particle reconstruction, and bioinformatics |
In progress |
Hanein (Gertler, Volkmann) |
| Formin-actin |
EM and biochemistry |
In progress |
Hanein (Higgs) |
| C-terminal fragment of Tensin and actin |
EM, helical reconstruction and bioinformatics |
In progress |
Hanein (Li, Li, Vuori, Volkmann, Liddington) |
| Talin-rod domain and actin |
EM and helical reconstruction |
In progress |
Hanein (Liddington) |
| Nanodisc |
Cryo EM |
In progress |
Hanein (Sligar, Volkmann) |
| Fimbrin-actin regulation |
EM and biochemistry |
In progress |
Hanein (Volkmann) |
| Fibronectin structure |
EM, Cryo EM and tomography |
In progress |
Hanein (Volkmann, Engel) |
| Full-length talin and actin |
EM and tomography |
In progress |
Hanein (Volkmann, Liddington) |
| Full-length Vinculin actin |
EM, tomography, single particle reconstructions, docking |
In progress |
Hanein (Volkmann, Liddington) |
| Full-length tensin and actin |
Cryo EM |
In progress |
Hanein (Vuori) |
| Calcium and integrin binding protein (CIB) complex with integrin |
X-Ray crystallography |
In progress |
Liddington |
| Calcium and integrin binding protein (CIB) complex with WASP |
X-Ray crystallography |
In progress |
Liddington |
| Paxillin-wt fragments 1-328 & 325-557 and full-length |
X-Ray crystallography |
In progress |
Liddington |
| Isolated membrane protrusions (with Klemke) |
Electron Tomography |
In progress |
Taylor |
| Crawling cells (with Condeelis) |
Electron Tomography |
In progress |
Taylor |
| Actin networks |
VMP, Cryo EM and tomography |
In progress |
Hanein (Volkmann) |
| Talin F3 domain with beta 3 integrin tail fused to PIPKIg complex |
NMR |
In progress - Assignment of F3 domain completed using NMR techniques-collaboratoin with Campbell and Ginsberg (Wegener et al. Cell 2007) |
Liddington |
| Vinculin and Talin |
Atomic Force Microscope |
In progress - Atomic Force Microscope experiments begun to monitor force required to stretch-activate viniculin and talin in vitro in presence of ligands, and correlate with cell based studies (Schwartz). |
Liddington |
| Podosome structure |
Correlative light microscopy-electron microscopy |
In progress - Collaboration established, ongoing research visit |
Geiger (Hanein) |
| Vinculin N-terminal domain complex with alpha-catenin domain 3 |
X-Ray crystallography |
In progress - Complex purified by gel filtration. Crystallization trials in progress |
Liddington |
| Metacinculin (fragments and full length) |
X-Ray crystallography, NMR |
In progress - crystalization trials. residues 879-1134; metavinculin residues 895-11; full-length metavinculin. All proteins expressed. |
Liddington |
| Talin-vinculin |
2D crystallization on lipid monolayers |
In progress - Crystallization trials underway |
Taylor |
| Talin |
2D crystallization on lipid monolayers |
In progress - Crystallization trials underway |
Taylor (Liddington) |
| Vinculin and Arp2/3 complex formation |
X-Ray crystallography |
In progress - crystallization trials. Vinculin residues 715-1066 (domains 4 and 5, including proline-rich region) expressed. Arp2/3 subunit p.34Arc human Arp_C2 domain (34.3kDa) and p20Arc: human Arp_C4 domain (19.5kDa) expressed stably. Complex formation in progress. |
Liddington |
| Alpha-actinin1 (in EGTA) |
2D crystallization on lipid monolayers |
In progress - Currently on hold pending expression and purification efforts |
Taylor |
| Alpha-actinin1 (plus calcium) |
2D crystallization on lipid monolayers |
In progress - Currently on hold pending expression and purification efforts |
Taylor |
| Paxillin (1-328)-FAT domain of FAK |
NMR |
In progress - FAT resonances assigned, various paxillin fragments expressed |
Campbell |
| Alpha-actinin1/F-actin complexes |
Electron Tomography |
In progress - Focal adhesion complexes on lipid monolayers. Assemby of complexes of F-actin with talin, full length vinculin, alpha-actin-1 and beta1-integrin cytoplasmic domain. Exploring effects of different lipid composition. |
Taylor |
| Layilin-talin (196-400) |
X-Ray crystallography, NMR |
In progress - Good expression but no crystals |
Liddington |
| Alpha-actinin beta1-integrin-vinculin1-258-talin FERM domain complex |
2D crystallization on lipid monolayers |
In progress - Large 2D arrays completed, 3D image underway. Identification of vinculin binding site on alpha-actinin; identification of beta1--integrin binding site on alpha-actinin. Nearing completion of ice embedded specimens |
Taylor (Liddington) |
| Organization of zyxin and alpha-actinin in focal adhesions and stress fibers |
Correlative light microscopy-electron microscopy |
In progress - Light microscopy aspect initiated |
Geiger (Hanein) |
| Mek1 |
X-Ray crystallography |
In progress - Protein expressed and purified from E. coli. Crystal trials in progress |
Liddington |
| Talin rod |
X-Ray crystallography |
In progress - Protein expressed and purified from E. coli. Crystal trials in progress. Complementary studies with Hanein in progress. |
Liddington |
| Talin (full length) |
X-Ray crystallography |
In progress - Protein expressed and purified from E.coli. Crystal trials in progress. Complementary EM studies with Hanein and Critchley in progress. |
Liddington |
| Talin fragments for protein splicing and chemical labeling |
Cell-based studies |
In progress - Protein expressed and purified from E.coli. Ligation in progress |
Liddington |
| Full-length FAK |
X-Ray crystallography |
In progress - Protein expressed and purified from insect cells. Crystal trials in progress |
Liddington |
| GIT-1 |
X-Ray crystallography |
In progress - Protein expressed and purified from insect cells. Crystal trials in progress |
Liddington |
| Beta 3-Src SH3 |
NMR |
In progress - Protein spectra assigned; currently being deuterated to study complex |
Campbell (Ginsberg) |
| Talin rod 655-911 |
NMR |
In progress - Resonance assignments of 8 helices ongoing |
Critchley |
| PIP3 binding DHR1 domain of DOCK180 |
X-Ray crystallography |
In progress - structural studies (with Kristiina Vuori and Jean-Francois Cote). Stable protein expression; crystalization trials in progress. |
Liddington |
| Beta 3-talin (306-406) |
NMR |
In progress - Structure of complex being refined |
Campbell (Ginsberg) |
| Talin FO/F1(1-200) |
NMR |
In progress - Structure of F0, F1, and F0F1 constructs have been solved. Model of the talin head based on the structure of F0F1 and F2F3 is being refined using SAXS data. Currently using NMR chemical shift experiments to map the Ha-Ras binding site in the head. |
Critchley |
| Talin C-terminal actin binding site (I/LWEQ motif) |
NMR |
In progress - Structure solved and submitted for publication |
Critchley |
| Talin C-terminal helix required for dimer formation |
X-Ray crystallography |
In progress - Structure solved to 2.5 Angstroms and submitted for publication |
Critchley |
| GIT-1 domains |
X-Ray crystallography |
In progress - Subcloning and expression trials in progress |
Liddington |
| Talin head for head-tail binding studies |
Calorimetric |
In progress - Talin head expressed in mg quantities. ITC and DSC studies in progress |
Liddington |
| alphaIIb-beta3 integrin structure in the membrane with Ligands and talin FERM domain |
Electron Tomography |
Under consideration |
Taylor |
| Refinement of docking algorithms for high-resolution interpretation of cryoEM maps |
Completed - Volkmann & Hanein, 2003 Shacham et al., 2007 |
Volkmann |
| Test and refinement of noise reduction algorithms for tomographic reconstructions |
Completed - Van der Heide et al., 2007 |
Volkmann (Hanein, Taylor, Marsh) |
| Use of reduced-representation templates for particle selection in cryoEM |
Completed - Volkmann, 2004 , Zhu et al. 2004 |
Volkmann |
| Application of docking algorithms for high-resolution interpretation of cryoEM maps |
Completed - Rouiller et al., 2008,Lowey et al., 2007,Trybus et al., 2007,Gingras et al., 2008,Janssen et al., 2006,Lee et al., 2004 |
Volkmann (Hanein, Liddington, Critchley) |
| Refinement of docking algorithms for use with tomograms |
Data available, first application published in Rouiller et al., 2008. Technical publication in preparation. Software available |
Volkmann (Hanein) |
| Refinement of reconstruction algorithms for helical structures |
Completed - Gingras et al., 2008,Lowey et al., 2007, Volkmann, et al., 2007, Trybus et al., 2007, Janssen et al., 2006, Craig et al., 2006. Initial version completed; more detailed studies in progress.Technical publication in preparation. |
Volkmann |
| Application of filament tracking approaches to interpretation of worm sperm tomograms |
In progress |
Volkmann (Taylor, Roberts) |
| Refinement of watershed segmentation for use with electron tomograms |
In progress - Initial test completed, publication: Volkmann, 2002 more detailed studies in progress |
Volkmann (Hanein, Taylor, Marsh) |
| Improved single-particle reconstruction algorithms for small particles and mixed populations |
In progress - initial tests completed |
Volkmann |
| Test and refinement of reduced-representation templates for filament detection in electron tomograms |
In progress - Initial tests completed, refinement of parameters in progress |
Volkmann (Hanein, Taylor) |
| Adaptation of watershed transform for use with light microscopy |
In progress - Initial tests successfully completed, more detailed studies in progress |
Volkmann (Hanein, Danuser, Waterman, Geiger) |
| Averaging of motifs extracted from tomographic reconstructions |
Initial version completed, first application published in Rouiller et al., 2008. Technical publication in preparation |
Volkmann |
