Decavanadate as a biochemical tool in the elucidation of muscle contraction regulation.
Recently reported decameric vanadate (V(10)) high affinity binding site in myosin S1, suggests that it can be used as a tool in the muscle contraction regulation. In the present article, it is shown that V(10) species induces myosin S1 cleavage, upon irradiation, at the 23 and 74 kDa sites, the latter being prevented by actin and the former blocked by the presence of ATP. Identical cleavage patterns were found for meta- and decavanadate solutions, indicating that V(10) and tetrameric vanadate (V(4)) have the same binding sites in myosin S1. Concentrations as low as 50 muM decavanadate (5 muM V(10) species) induces 30% of protein cleavage, whereas 500 muM metavanadate is needed to attain the same extent of cleavage. After irradiation, V(10) species is rapidly decomposed, upon protein addition, forming vanadyl (V(4+)) species during the process. It was also observed by NMR line broadening experiments that, V(10) competes with V(4) for the myosin S1 binding sites, having a higher affinity. In addition, V(4) interaction with myosin S1 is highly affected by the products release during ATP hydrolysis in the presence or absence of actin, whereas V(10) appears to be affected at a much lower extent. From these results it is proposed that the binding of vanadate oligomers to myosin S1 at the phosphate loop (23 kDa site) is probably the cause of the actin stimulated myosin ATPase inhibition by the prevention of ATP/ADP exchange, and that this interaction is favoured for higher vanadate anions, such as V(10).
Tiago
Teresa
T
Departamento de Química e Bioquímica, Faculdade de Ciências e Tecnologia, Universidade do Algarve, FCT, UALG, Gambelas, 8005-139 Faro, Portugal. ttiago@ualg.pt
Aureliano
Manuel
M
Moura
José J G
JJ
D000818:Animals; D066298:In Vitro Techniques; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D009119:Muscle Contraction / Q000187:drug effects* / Q000502:physiology / Q000528:radiation effects; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000502:physiology / Q000528:radiation effects; D011817:Rabbits; D013053:Spectrophotometry; D014638:Vanadates / Q000494:pharmacology*
D014638:Vanadates