Counteracting effects of urea and methylamines in function and structure of skeletal muscle myosin.
Myosin is an asymmetric protein that comprises two globular heads (S1) and a double-stranded alpha-helical rod. We have investigated the effects of urea and the methylamines trimethylamine oxide (TMA-O) and glycine betaine (betaine) on activity and structure of skeletal muscle myosin. K(+) EDTA ATPase activity of myosin was almost completely inhibited by urea (2M); TMA-O stimulated myosin activity, whereas betaine had no effect. When combined with urea (0-2M), TMA-O or betaine (1 M) effectively protected the ATPase activity of myosin against inhibition. Intrinsic fluorescence measurements showed that in urea or TMA-O (0-2M), there were no shifts in the center of mass of the fluorescence spectrum of myosin, despite a decrease in fluorescence intensity. However, these osmolytes at concentrations above 2M produced a red shift in the emission spectrum. Betaine alone did not alter the center of mass at any concentration tested up to 5.2M. Thus, modifications in ATPase activity induced by low concentrations of solutes (<2M) are not directly correlated with the modifications in myosin structure detected by fluorescence. Both methylamines (>or=1M) were also able to protect myosin structure against urea-induced effects (2-8M). Protection was not observed for S1, supporting the hypothesis that these osmolytes have a biphasic effect on myosin: at lower concentrations there is an effect on the globular portion (S1), and at higher concentrations there is an effect on the coiled-coil (rod) portion of myosin.
Ortiz-Costa
Susana
S
Laboratório de Enzimologia e Controle do Metabolismo (LabECoM), Departamento de Fármacos, Faculdade de Farmácia/CCS/UFRJ, Rio de Janeiro, Brazil.
Sorenson
M M
MM
Sola-Penna
Mauro
M
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001622:Betaine / Q000494:pharmacology; D005453:Fluorescence; D008744:Methylamines / Q000494:pharmacology*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D024744:Skeletal Muscle Myosins / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D014508:Urea / Q000494:pharmacology*
D008744:Methylamines; D001622:Betaine; D014508:Urea; D000251:Adenosine Triphosphatases; D024744:Skeletal Muscle Myosins; C005855:trimethyloxamine