Drug action of ritodrine on the sarcoplasmic reticulum Ca(2+)-ATPase from skeletal muscle.
Ritodrine inhibits the steady-state Ca(2+)-ATPase activity of isolated sarcoplasmic reticulum vesicles in a dose-dependent manner. The observed K0.5 value for inhibition (approximately 3 mM) gives proof of a low-affinity interaction. Ritodrine also interferes with the steady-state Ca2+ transport ability decreasing the maximal rate without modification of the Ca2+ or ATP affinity for the enzyme. This is consistent with an absence of competition for the transport and the catalytic sites. Analysis of the catalytic and transport cycle shows that: (i) ritodrine inhibits the phosphorylation partial reaction. This is supported by pre-steady-state kinetic experiments of Ca2+ transport and also by the temperature dependence of the phosphoenzyme level. (ii) At high ritodrine concentrations the dephosphorylation step becomes rate-limiting. This is suggested by the biphasic profile (V-shape) of phosphoenzyme accumulation at different ritodrine concentrations. This was also confirmed by chase experiments of radioactive phosphoenzyme decomposition at steady state. These data reveal a complex pattern of inhibition involving two sites for interaction with low and high ritodrine concentrations. It is envisaged that ritodrine does not exert any direct effect on the smooth muscle sarcoplasmic reticulum Ca(2+)-ATPase when used in the treatment of preterm birth.
Caravaca
F
F
Departamento de Bioquimica y Biologia Molecular, Edificio de Veterinaria, Universidad de Murcia, Espinardo, Spain.
Soler
F
F
Gomez-Fernandez
J C
JC
Fernandez-Belda
F
F
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D017136:Ion Transport; D007700:Kinetics; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000201:enzymology*; D007752:Obstetric Labor, Premature / Q000517:prevention & control; D010766:Phosphorylation; D011247:Pregnancy; D011817:Rabbits; D012312:Ritodrine / Q000737:chemistry / Q000494:pharmacology*; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000201:enzymology; D013329:Structure-Activity Relationship; D013696:Temperature
D000255:Adenosine Triphosphate; D000252:Calcium-Transporting ATPases; D012312:Ritodrine; D002118:Calcium