Molecular basis of myosin-2 function

• Applicant: Privatdozent Dr. Walter Steffen
• Subject Area: Cell Biology
• Term: from 2009 to 2013
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 14023866

Myosin-2 isoforms vary widely in terms of their mechanical and biochemical properties thereby in-fluencing their functionality. The aim of this project is to elucidate the molecular basis for the different functions at the single molecule level. Almost all our knowledge about the mechanochemical interac-tions of actomyosin is derived from averaged data sets; either from solution studies to determine kinetic parameters of the actomyosin interactions or from investigations of the motor activity in a filament gliding assay using an ensemble of motor proteins. Averaging methods are also used in single molecule studies. As a result of averaging procedures a load dependent bias can not be excluded. By following a single molecule of slow non-muscle myosins through the complete actomyosin cross-bridge cycle, we will dissect the biochemical and mechanical steps of the cycle from the initial weak binding state to the rigor state. Combining an improved time resolution optical trap with total internal reflection (TIRF) microscopy to assess nucleotide state will allow us to study the coupling of biochemical and molecular events at the single molecule level.

• DFG Programme: Research Units
• Subproject of: FOR 629: Molecular Mechanisms of Cell Motility