title: An actin-dependent conformational change in myosin.
abstract: Conformational changes within myosin lead to its movement relative to an actin filament. Several crystal structures exist for myosin bound to various nucleotides, but none with bound actin. Therefore, the effect of actin on the structure of myosin is poorly understood. Here we show that the swing of smooth muscle myosin lever arm requires both ADP and actin. This is the first direct observation that a conformation of myosin is dependent on actin. Conformational changes within myosin were monitored using fluorescence resonance energy transfer techniques. A cysteine-reactive probe is site-specifically labeled on a 'cysteine-light' myosin variant, in which the native reactive cysteines were removed and a cysteine engineered at a desired position. Using this construct, we show that the actin-dependent ADP swing causes an 18 A change in distance between a probe on the 25/50 kDa loop on the catalytic domain and a probe on the regulatory light chain, corresponding to a 23 degrees swing of the light-chain domain.
authors.affiliation: Department of Physics, University of Illinois, 1110 West Green Street, Urbana, Illinois 61801, USA.
authors.forename: Ming
authors.initials: M
authors.lastname: Xiao
authors.forename: Jeff G
authors.initials: JG
authors.lastname: Reifenberger
authors.forename: Amber L
authors.initials: AL
authors.lastname: Wells
authors.forename: Corry
authors.initials: C
authors.lastname: Baldacchino
authors.forename: Li-Qiong
authors.initials: LQ
authors.lastname: Chen
authors.forename: Pinghua
authors.initials: P
authors.lastname: Ge
authors.forename: H Lee
authors.initials: HL
authors.lastname: Sweeney
authors.forename: Paul R
authors.initials: PR
authors.lastname: Selvin
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000378:metabolism*; D019943:Amino Acid Substitution; D000818:Animals; D002645:Chickens; D003545:Cysteine; D031541:Fluorescence Resonance Energy Transfer; D005895:Gizzard, Avian / Q000378:metabolism; D007700:Kinetics; D009130:Muscle, Smooth / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D018994:Myosin Light Chains / Q000737:chemistry / Q000378:metabolism; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D000199:Actins; D018994:Myosin Light Chains; D009218:Myosins; D003545:Cysteine