title: MgATP-induced conformational changes in a single myosin molecule observed by atomic force microscopy: periodicity of substructures in myosin rods.
abstract: This paper discusses the conformational changes in a single myosin molecule directly observed using atomic force microscopy (AFM). The myosin molecules were pretreated in rigor solutions without MgATP or in relaxed solutions with various concentrations of MgATP. The images of these molecules were obtained using a tapping mode AFM. The results indicate that the orientation of the myosin's heads and tail strongly depend on the MgATP concentration. Without using MgATP, almost all of the myosin molecules are in the extended form; however, when MgATP is used, the molecules bend according to the level of MgATP concentration. The mean-square end-to-end distance of the myosin molecules is significantly shorter with p[MgATP] = 4 than with p[MgATP] = 6. The rod region did not show the same level of intensity along their length in the extended form. The rods exhibited clusters of discontinuity, which were identified as substructures. The size of these substructures change at intervals that are multiples of 14.3-14.5 nm, which reflects the periodicity of the alpha-helical coiled coils. The substructure clusters also correspond to the myosin crossbridge spacing in muscles (14.3 or 43 nm). These results suggest that the myosin's head bends in conjunction with the bending or tilting in the helical substructures. Conformational changes of the myosin molecule induced by MgATP seem to mimic the molecular motions in a muscle's force generation process.
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Nagoya University, Nagoya, Japan. mtanig@nubio.nagoya-u.ac.jp
authors.forename: Mieko
authors.initials: M
authors.lastname: Taniguchi
authors.forename: Osamu
authors.initials: O
authors.lastname: Matsumoto
authors.forename: Shigehito
authors.initials: S
authors.lastname: Suzuki
authors.forename: Yasunobu
authors.initials: Y
authors.lastname: Nishino
authors.forename: Ayumu
authors.initials: A
authors.lastname: Okuda
authors.forename: Tooru
authors.initials: T
authors.lastname: Taga
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamane
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D018625:Microscopy, Atomic Force / Q000379:methods; D018482:Muscle, Skeletal / Q000737:chemistry / Q000648:ultrastructure*; D015879:Myosin Subfragments / Q000737:chemistry / Q000648:ultrastructure; D009218:Myosins / Q000737:chemistry* / Q000187:drug effects / Q000648:ultrastructure*; D011487:Protein Conformation; D011817:Rabbits
chemical_list: D015879:Myosin Subfragments; D000255:Adenosine Triphosphate; D009218:Myosins