Correlation of ActoS1, myofibrillar, and muscle fiber ATPases.
Our objective was to determine a good in vitro model for muscle fiber ATPase, and we compared the kinetics of Ca(2+)-activated myofibrils and cross-linked actoS1 in a buffer of physiological ionic strength. The myofibrils were cross-linked chemically to mimic the isometric condition of fibers or were un-cross-linked (the isotonic condition), and temperature perturbation was used to probe their ATPase mechanisms. At 4 degrees C, we have already shown that the kinetics of cross-linked actoS1 and myofibrils (cross-linked or not) are similar: there were large P(i) bursts and kcat values of about 1 s-1, close to that obtained with fibers [Herrmann, C., Sleep, J., Chaussepied, P., Travers, F. & Barman, T. (1993) Biochemistry 32, 7255-7263]. So, at 4 degrees C cross-linked actoS1 and myofibrils are equally good as models for fiber ATPase. At 20 degrees C, this similarity vanishes: progress curves with the myofibrils (cross-linked or not) had large P(i) bursts, but with cross-linked actoS1, bursts could not be discerned. This shows that at 20 degrees C the predominant steady-state intermediates are ATP complexes with actoS1 but are products complexes with the myofibrils, as with fibers [Ferenczi, M.A. (1986) Biophys. J. 50, 471-477]. Further, the kcat values were different: 15.5 s-1 with cross-linked actoS1, 8.3 s-1 for myofibrils, and 3.5 s-1 for cross-linked myofibrils. With fibers, kcat = 3.3 s-1. These results show that cross-linked myofibrillar ATPase is a good model for muscle fibers contracting isometrically.(ABSTRACT TRUNCATED AT 250 WORDS)
Herrmann
C
C
INSERM U128, CNRS, BP 5051, Montpellier, France.
Lionne
C
C
Travers
F
F
Barman
T
T
D000205:Actomyosin / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D008954:Models, Biological; D009132:Muscles / Q000201:enzymology*; D013696:Temperature
D000255:Adenosine Triphosphate; D000205:Actomyosin; D000251:Adenosine Triphosphatases; D002118:Calcium