title: Engineering a light-controlled F1 ATPase using structure-based protein design.
abstract: The F1 sub-complex of ATP synthase is a biological nanomotor that converts the free energy of ATP hydrolysis into mechanical work with an astonishing efficiency of up to 100% (Kinosita et al., 2000). To probe the principal mechanics of the machine, I re-engineered the active site of E.coli F1 ATPase with a structure-based protein design approach: by incorporation of a site-specific, photoswitchable crosslinker, whose end-to-end distance can be modulated by illumination with light of two different wavelengths, a dynamic constraint was imposed on the inter-atomic distances of the α and β subunits. Crosslinking reduced the ATP hydrolysis activity of four designs tested in vitro and in one case created a synthetic ATPase whose activity can be reversibly modulated by subsequent illumination with near UV and blue light. The work is a first step into the direction of the long-term goal to design nanoscaled machines based on biological parts that can be precisely controlled by light.
authors.affiliation: Experimental Molecular Biophysics, Department of Physics, Freie Universität Berlin , Berlin , Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Hoersch
keywords: Azobenzene; Light control; Molecular machine; Protein design