Sliding velocity of isolated rabbit cardiac myosin correlates with isozyme distribution.
To investigate the relationship between the mechanical and biochemical properties of cardiac myosin, the sliding velocity of isolated cardiac myosin obtained from both euthyroid and hyperthyroid rabbits on actin cables was measured with an in vitro motility assay system. Ten rabbits (T) were treated with L-thyroxine to induce hyperthyroidism, and eight nontreated animals (N) were used as controls. Myosin was purified from the left ventricles of anesthetized animals. Myosin isozyme content was analyzed by the pyrophosphate gel electrophoresis method, and myosin adenosinetriphosphatase (ATPase) activity was determined on the same sample. Long well-organized actin cables of green algae, Nitellopsis, were used in the in vitro motility assay. Small latex beads were coated with purified cardiac myosin and introduced onto the Nitellopsis actin cables. Active unidirectional movement of the beads on the actin cables was observed under a photomicroscope, and the velocity was measured. The velocity was dependent on ATP concentrations, and the optimal pH for bead movement was approximately 7.0-7.5. The mean velocity was higher in T than in N (0.66 +/- 0.12 vs. 0.32 +/- 0.09 micron/s, P less than 0.01). Both Ca(2+)-activated ATPase activity and the percentage of alpha-myosin heavy chain were also higher in T than in N (0.691 +/- 0.072 vs. 0.335 +/- 0.072 microM Pi.mg-1.min-1, P less than 0.01, and 79 +/- 12 vs. 26 +/- 7%, P less than 0.01, respectively). The velocity of myosin closely correlated with both Ca(+2)-activated myosin ATPase activity (r = 0.87, P less than 0.01) and the percentage of alpha-myosin heavy chain (r = 0.87, P less than 0.01).
Yamashita
H
H
Second Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Sugiura
S
S
Serizawa
T
T
Sugimoto
T
T
Iizuka
M
M
Katayama
E
E
Shimmen
T
T
D000818:Animals; D005260:Female; D006980:Hyperthyroidism / Q000201:enzymology / Q000473:pathology; D007527:Isoenzymes / Q000302:isolation & purification / Q000502:physiology*; D007840:Latex; D008297:Male; D008854:Microscopy, Electron; D008863:Microspheres; D009206:Myocardium / Q000201:enzymology* / Q000648:ultrastructure; D009218:Myosins / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D011817:Rabbits; D012016:Reference Values
D007527:Isoenzymes; D007840:Latex; D009218:Myosins