Beta-tubulin and benzimidazole resistance in the sheep nematode Haemonchus contortus

Abstract

We have compared benzimidazole (BZ) susceptible ( S) and resistant ( R) strains of Haemonchus contortus from sheep by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), Western blotting and ELISA techniques. The S strain bound more drug per mg protein than the R strain. BZ binding could be resolved into high-affinity and low-affinity binding. Low-affinity binding in parasite preparations devoid of tubulin was observed, but high-affinity binding occurred only in preparations containing tubulin. Resistance was associated with a decrease in the high affinity component. The S and R strains were shown by ELISA to contain similar total amounts of tubulin. By 2-D PAGE, the β-tubulin isoform pattern of the S strain was different from that of the R strain, but the α-tubulin isoform patterns of the 2 strains were similar. BZ resistance was associated with a decrease in high-affinity BZ binding to tubulin and an alteration in β-tubulin isoform pattern.