The two actin-binding regions on the myosin heads of cardiac muscle.
In the presence of myosin S1 or myosin heads, actin filaments tend to form bundles. The biological meaning of the bundling of actin filaments has been unclear. In this study, we found that the cardiac myosin heads can form the bundles of actin filaments more rapidly than can skeletal S1, as monitored by light scattering and electron microscopy. Moreover, the actin bundles formed by cardiac S1 were found to be more stable against mechanical agitation. The distance between actin filaments in the bundles was approximately 20 nm, which is comparable to the length of a myosin head and two actin molecules. This suggests the direct binding of S1 tails to the adjacent actin filament. The "essential" light chain of cardiac myosin could be cross-linked to the actin molecule in the bundle. When monomeric actin molecules were added to the bundle, the bundles could be dispersed into individual filaments. The three-dimensional structure of the dispersed actin filaments was reconstructed from electron cryo-microscopic images of the single actin filaments dispersed by monomer actin. We were able to demonstrate that cardiac myosin heads bind to two actin molecules: one actin molecule at the conventional actin-binding region and the other at the essential light-chain-binding region. This capability of cardiac myosin heads to bind two actin molecules is discussed in view of lower ATPase activity and slower shortening velocity than those of skeletal ones.
Miyanishi
Takayuki
T
Faculty of Environmental Studies, Nagasaki University, Nagasaki 852-8521, Japan. miyanish@net.nagasaki-u.ac.jp
Ishikawa
Takashi
T
Hayashibara
Toshihisa
T
Maita
Tetsuo
T
Wakabayashi
Takeyuki
T
D000199:Actins / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens; D003432:Cross-Linking Reagents / Q000378:metabolism; D008840:Microfilament Proteins / Q000378:metabolism* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000648:ultrastructure; D009206:Myocardium / Q000378:metabolism* / Q000648:ultrastructure; D018994:Myosin Light Chains / Q000378:metabolism* / Q000648:ultrastructure; D015879:Myosin Subfragments / Q000378:metabolism / Q000648:ultrastructure; D011485:Protein Binding; D012542:Scattering, Radiation; D013388:Succinimides / Q000378:metabolism
D000199:Actins; D003432:Cross-Linking Reagents; D008840:Microfilament Proteins; D018994:Myosin Light Chains; D015879:Myosin Subfragments; D013388:Succinimides; C066969:maleimidobenzoyl-G-actin; C013097:3-maleimidobenzoyl N-hydroxysuccinimide; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate