Decavanadate interactions with actin: inhibition of G-actin polymerization and stabilization of decameric vanadate.
Decameric vanadate species (V10) inhibit the rate and the extent of G-actin polymerization with an IC50 of 68+/-22 microM and 17+/-2 microM, respectively, whilst they induce F-actin depolymerization at a lower extent. On contrary, no effect on actin polymerization and depolymerization was detected for 2mM concentration of "metavanadate" solution that contains ortho and metavanadate species, as observed by combining kinetic with (51)V NMR spectroscopy studies. Although at 25 degrees C, decameric vanadate (10 microM) is unstable in the assay medium, and decomposes following a first-order kinetic, in the presence of G-actin (up to 8 microM), the half-life increases 5-fold (from 5 to 27 h). However, the addition of ATP (0.2mM) in the medium not only prevents the inhibition of G-actin polymerization by V10 but it also decreases the half-life of decomposition of decameric vanadate species from 27 to 10h. Decameric vanadate is also stabilized by the sarcoplasmic reticulum vesicles, which raise the half-life time from 5 to 18h whereas no effects were observed in the presence of phosphatidylcholine liposomes, myosin or G-actin alone. It is proposed that the "decavanadate" interaction with G-actin, favored by the G-actin polymerization, stabilizes decameric vanadate species and induces inhibition of G-actin polymerization. Decameric vanadate stabilization by cytoskeletal and transmembrane proteins can account, at least in part, for decavanadate toxicity reported in the evaluation of vanadium (V) effects in biological systems.
Ramos
Susana
S
Dept. Química e Bioquímica, FCT, Universidade do Algarve, Faro, Portugal.
Manuel
Miguel
M
Tiago
Teresa
T
Duarte
Rui
R
Martins
Jorge
J
Gutiérrez-Merino
Carlos
C
Moura
José J G
JJ
Aureliano
Manuel
M
D000199:Actins / Q000737:chemistry* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009218:Myosins / Q000737:chemistry / Q000378:metabolism; D011108:Polymers / Q000737:chemistry; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D014638:Vanadates / Q000737:chemistry* / Q000378:metabolism
D000199:Actins; D008081:Liposomes; D011108:Polymers; D014638:Vanadates; D000255:Adenosine Triphosphate; D009218:Myosins