Preliminary characterization and interaction of tubulin from Trichinella spiralis larvae with benzimidazole derivatives.
Tubulin was estimated to account for 0.3% of the total soluble protein in Trichinella spiralis cytosolic fractions. Tubulin from T. spiralis was partially purified by precipitation with either taxol or vinblastine sulphate. Immunoblotting with alpha- and beta-tubulin monoclonal antibodies revealed the presence of tubulin in T. spiralis partially purified preparations. Electrophoretic mobility of T. spiralis tubulin in sodium dodecyl sulphate-polyacrylamide gels was very similar to that shown by pig brain tubulin. Further studies with colchicine binding assays indicated that T. spiralis tubulin has binding features similar to that of tubulin from other nematodes: colchicine association constant = 8.1 x 10(-4) M and competitive inhibition of colchicine binding by podophyllotoxin, with an inhibition constant of 1.3 x 10(-6) M. Finally, inhibition of colchicine binding by several benzimidazoles (mebendazole, fenbendazole, oxibendazole and albendazole) was investigated. All the benzimidazoles inhibited colchicine binding in a competitive manner, with inhibition constant values ranging from 1.4 x 10(-7) M (mebendazole) to 3.9 x 10(-6) M (fenbendazole).
Jiménez-González
A
A
Parasitology Laboratory, Faculty of Pharmacy, University of Alcaldá de Henares, Madrid, Spain.
De Armas-Serra
C
C
Criado-Fornelio
A
A
Casado-Escribano
N
N
Rodríguez-Caabeiro
F
F
Díez
J C
JC
D015766:Albendazole / Q000378:metabolism; D000818:Animals; D000871:Anthelmintics / Q000378:metabolism; D001562:Benzimidazoles / Q000378:metabolism*; D001667:Binding, Competitive; D003078:Colchicine / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005273:Fenbendazole / Q000378:metabolism; D015151:Immunoblotting; D008463:Mebendazole / Q000378:metabolism; D014234:Trichinella / Q000032:analysis*; D014404:Tubulin / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D000871:Anthelmintics; D001562:Benzimidazoles; D014404:Tubulin; C011321:oxibendazole; D005273:Fenbendazole; D008463:Mebendazole; D015766:Albendazole; D003078:Colchicine