This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We performed Small Angle X-ray Scattering (SAXS) structural studies using the BioCAT beamline on a novel actin filament nucleator from Vibrio parahaemolyticus named VopL. We have found that the strong actin filament nucleation activity of VopL depends on the presence of three WH2 domains and dimerization through a C-terminal domain (DD) of unknown structure. Since this DD had no detectable similarity to any known structure in the Protein Data Bank, we used SAXS to study its structure in solution to gain further insights into the nucleation mechanism of VopL. The SAXS structure showed the expected two-fold symmetry of the dimer, consisting of a central dimerization 'knot domain', which probably comprises the predicted C-terminal coiled coil, and two diametrically opposed smaller domains. We also studied the complex of actin with VopL fragment 1W-DD (containing one of the three WH2 domains and the DD).. While the study of this complex did not yield much new information, a few inferences may be drawn: a) the structure of the DD reemerged as a stable unit in the structure of the complex with actin, b) actin subunits are bound loosely in the polymerization nucleus and c) the actin subunits bind at the interface between the central knob and small distal domains. In summary, our biochemical and structural analyses by SAXS of VopL constructs ?actin suggest that the dimerization domain binds at the pointed end of the actin filament and contains a coiled-coil region at the C-terminus.

National Institute of Health (NIH)
National Center for Research Resources (NCRR)
Biotechnology Resource Grants (P41)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-BCMB-E (40))
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Illinois Institute of Technology
Other Basic Sciences
Schools of Arts and Sciences
United States
Zip Code
Yazdi, Aliakbar Khalili; Vezina, Grant C; Shilton, Brian H (2017) An alternate mode of oligomerization for E. coli SecA. Sci Rep 7:11747
Sullivan, Brendan; Robison, Gregory; Pushkar, Yulia et al. (2017) Copper accumulation in rodent brain astrocytes: A species difference. J Trace Elem Med Biol 39:6-13
Morris, Martha Clare (2016) Nutrition and risk of dementia: overview and methodological issues. Ann N Y Acad Sci 1367:31-7
Robison, Gregory; Sullivan, Brendan; Cannon, Jason R et al. (2015) Identification of dopaminergic neurons of the substantia nigra pars compacta as a target of manganese accumulation. Metallomics 7:748-55
Gelfand, Paul; Smith, Randy J; Stavitski, Eli et al. (2015) Characterization of Protein Structural Changes in Living Cells Using Time-Lapsed FTIR Imaging. Anal Chem 87:6025-31
Liang, Wenguang G; Ren, Min; Zhao, Fan et al. (2015) Structures of human CCL18, CCL3, and CCL4 reveal molecular determinants for quaternary structures and sensitivity to insulin-degrading enzyme. J Mol Biol 427:1345-1358
Zhou, Hao; Li, Shangyang; Badger, John et al. (2015) Modulation of HIV protease flexibility by the T80N mutation. Proteins 83:1929-39
Nobrega, R Paul; Arora, Karunesh; Kathuria, Sagar V et al. (2014) Modulation of frustration in folding by sequence permutation. Proc Natl Acad Sci U S A 111:10562-7
Jiao, Lianying; Ouyang, Songying; Shaw, Neil et al. (2014) Mechanism of the Rpn13-induced activation of Uch37. Protein Cell 5:616-30
Witayavanitkul, Namthip; Ait Mou, Younss; Kuster, Diederik W D et al. (2014) Myocardial infarction-induced N-terminal fragment of cardiac myosin-binding protein C (cMyBP-C) impairs myofilament function in human myocardium. J Biol Chem 289:8818-27

Showing the most recent 10 out of 96 publications