title: Molecular engineering of a backwards-moving myosin motor.
abstract: All members of the diverse myosin superfamily have a highly conserved globular motor domain that contains the actin- and nucleotide-binding sites and produces force and movement. The light-chain-binding domain connects the motor domain to a variety of functionally specialized tail domains and amplifies small structural changes in the motor domain through rotation of a lever arm. Myosins move on polarized actin filaments either forwards to the barbed (+) or backwards to the pointed (-) end. Here, we describe the engineering of an artificial backwards-moving myosin from three pre-existing molecular building blocks. These blocks are: a forward-moving class I myosin motor domain, a directional inverter formed by a four-helix bundle segment of human guanylate-binding protein-1 and an artificial lever arm formed by two alpha-actinin repeats. Our results prove that reverse-direction movement of myosins can be achieved simply by rotating the direction of the lever arm 180 degrees.
authors.affiliation: Institut für Biophysikalische Chemie, Medizinische Hochschule Hannover, OE 4350, Carl-Neuberg-Strasse 1, D-30623 Hannover, Germany.
authors.forename: Georgios
authors.initials: G
authors.lastname: Tsiavaliaris
authors.forename: Setsuko
authors.initials: S
authors.lastname: Fujita-Becker
authors.forename: Dietmar J
authors.initials: DJ
authors.lastname: Manstein
mesh_terms: D000185:Actinin / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000818:Animals; D001696:Biomechanical Phenomena; D004268:DNA-Binding Proteins / Q000378:metabolism; D004023:Dictyostelium / Q000737:chemistry; D019204:GTP-Binding Proteins / Q000378:metabolism; D008954:Models, Biological; D008958:Models, Molecular; D020409:Molecular Motor Proteins / Q000737:chemistry / Q000378:metabolism*; D009218:Myosins / Q000737:chemistry / Q000378:metabolism*; D015202:Protein Engineering; D017434:Protein Structure, Tertiary
chemical_list: D000199:Actins; D004268:DNA-Binding Proteins; C490287:GBP1 protein, human; D020409:Molecular Motor Proteins; D000185:Actinin; D019204:GTP-Binding Proteins; D009218:Myosins