Association of Energy-Metabolism Enzymes with the Switch-Motor Complex of Bacterial Flagella

Project: Research project

Project Details

Description

Association of Energy-Metabolism Enzymes with the Switch-Motor Complex of Bacterial Flagella Association of Energy-Metabolism Enzymes with the Switch-Motor Complex of Bacterial Flagella. Our earlier studies demonstrated that fumarate reductase (FRD) forms a dynamic complex with the flagellar switch protein FliG of Escherichia coli and suggested that additional energy-metabolism enzymes, including the F1Fo ATP synthase, may be associated with the flagellar switch-motor complex, forming a supercomplex. Our aims in this proposal are to determine whether additional energy-metabolism enzymes are associated with the switch-motor complex, to identify them and the functions that they fulfill within the supercomplex, and to reveal structural information as to how an interaction between rather bulky membrane-linked complexes does not sterically interfere with flagellar rotation. We will screen for proteins interacting with the switch-motor complex both phylogenetically looking for proteins that have phylogenetic profiles similar to those of the switch proteins, and biochemically by identifying proteins in cell lysates that can be pulled-down with switch proteins. We will verify the binding in vitro by surface-plasmon resonance (SPR), analytical ultracentrifugation, and immuno-electron microscopy, and in vivo by bimolecular fluorescence complementation (BIFC). We will examine the effects of the interactions on the functions of the interacting proteins by biochemical and other approaches including single-molecule measurements of the effect of switch proteins on the F1Fo rotary motor. Last but not least, we will try revealing the molecular detail of the FRD-FliG interaction by crystallizing the FRD-FliG complex. Successful accomplishment of the proposed study may provide a clue to the puzzling question of why energy-metabolism systems are required near the switch-motor complex, and it is expected to reveal new insights into the overall control of flagellar function by interactions with proteins of otherwise diverse function.
StatusFinished
Effective start/end date9/1/108/31/14

Funding

  • United States-Israel Binational Science Foundation: $31,560.00

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