Studies proposed in this competing renewal application are intended to dissect how tensile mechanical force increases ability of some of the most common bacterial adhesins to bind carbohydrate ligands, i.e. mediate catch-bond mechanism of receptor-ligand interaction. Our current Bioengineering Research Partnership grant project has demonstrated that the strength of mannose-specific, FimH-mediated adhesion of Escherichia coli is dramatically enhanced by the presence of shear stress. We have shown that FimH adhesin is an allosterically regulated protein, where induced-fit mechanism of interaction of the mannose ligand with the FimH binding pocket is conformationally linked with separation between mannose-binding and fimbria-incorporating domains of FimH - the configuration favored by tensile mechanical force. In the renewal application, we propose to use, among other approaches, nuclear magnetic resonance (NMR) spectroscopy and atomic force microscopy (AFM) and molecular dynamics simulation (MD/SMD) to derive a comprehensive understanding of the conformational shift in course of FimH activation by the ligand and facilitation of this process by tensile force. We will use this knowledge for developing strategies on preventing adhesion of medically-relevant bacteria to host target cells and surfaces, for developing shear- modulated nanotechnological tools, and as a paradigm for understanding other types of shear-dependent bacterial adhesion.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI050940-09
Application #
8099765
Study Section
Special Emphasis Panel (ZRG1-IDM-H (02))
Program Officer
Baqar, Shahida
Project Start
2001-12-01
Project End
2013-06-30
Budget Start
2011-07-01
Budget End
2013-06-30
Support Year
9
Fiscal Year
2011
Total Cost
$478,061
Indirect Cost
Name
University of Washington
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
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Tchesnokova, Veronika; Aprikian, Pavel; Kisiela, Dagmara et al. (2011) Type 1 fimbrial adhesin FimH elicits an immune response that enhances cell adhesion of Escherichia coli. Infect Immun 79:3895-904
Pereverzev, Yuriy V; Prezhdo, Eugenia; Sokurenko, Evgeni V (2011) The two-pathway model of the biological catch-bond as a limit of the allosteric model. Biophys J 101:2026-36
Aprikian, Pavel; Interlandi, Gianluca; Kidd, Brian A et al. (2011) The bacterial fimbrial tip acts as a mechanical force sensor. PLoS Biol 9:e1000617

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