Structure transition in myosin association with the change of concentration: solubility equilibrium under specified KCl and pH condition.
We observed, for the first time, the elementary process for the ordered self-assembly formation of myosin in solution. It was realized exclusively under the specific condition of 200 mM KCl, 5 mM phosphate buffer, pH 7.08, at 15-20 degrees C, which is called the transition-generating condition (TGC). Described more in detail: pure myosin extracted from rabbit skeletal muscle exhibited the structural transition in its association form only when the myosin concentration c was changed under TGC. The myosin solubility was saturated in both edges of the total myosin concentration c > 10.0 mg/mL (solubility region II) and c < or = 0.25 mg/mL (solubility region I). In the intermediate region, the association structure of myosin changed stepwise with decreasing c. The steps were classified into four regions: region I (c < or = 0.25 mg/mL), II (0.25 < or = c < or = 0.50 mg/mL), III (0.50 < or = c < or = 5.0 mg/mL), and IV (c > 5.0 mg/mL). In each region except II, the plot of the relative soluble myosin concentration c(aq)/c against c(-1) gave a straight line of different slopes, certifying that myosin constructs self-assemblies by the closed association mechanism and that the self-assembly takes dual structures in each region. In region II, a drastic transition occurred in the self-assembled dual structures. Here, a highly associated (insoluble) giant assembly would break into soluble assemblies composed of several myosin molecules. The solubility region I originates a driving force for this structural transition. The basic binding unit of the self-assembly would be a parallel myosin-dimer constructed by the intermolecular axial staggers of 14.3 and 43 nm, as is observed by X-ray diffraction for the thick filament assembly or light meromyosin paracrystals. Myosin could take a single rod-like chain form only in an extremely low concentration region of c < or = c(aq,0) (= 0.053 mg/mL). The association behavior revealed in the present study suggests strongly that the complementary charge cluster and its electrostatic interaction between parallel myosin rods play a crucial role for the ordered self-assembly formation and that the specific electrostatic atmosphere of the solution under TGC is essential to the association mechanism in skeletal muscle myosin, or the thick filament formation of the mammals.
Tsunashima
Yoshisuke
Y
Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan. yh-tsunashima@r7.dion.ne.jp
Akutagawa
Tohru
T
D000818:Animals; D019281:Dimerization; D006863:Hydrogen-Ion Concentration / Q000187:drug effects*; D007526:Isoelectric Point; D018482:Muscle, Skeletal / Q000737:chemistry; D015879:Myosin Subfragments / Q000737:chemistry; D009218:Myosins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D009994:Osmolar Concentration; D011189:Potassium Chloride / Q000494:pharmacology*; D011817:Rabbits; D012995:Solubility / Q000187:drug effects*; D055672:Static Electricity; D014867:Water / Q000737:chemistry
D015879:Myosin Subfragments; D014867:Water; D011189:Potassium Chloride; D009218:Myosins