Molecular characterisation and localisation of an Onchocerca volvulus pi-class glutathione S-transferase.
Glutathione S-transferases (GSTs) constitute a major detoxification mechanism in helminth organisms and are regarded vaccine candidates against helminth infections. Onchocerca volvulus glutathione-binding proteins were purified from the aqueous soluble fraction of homogenised adult females by affinity chromatography on glutathione-agarose. The eluted proteins had a specific GST activity of 1.6 mumol min-1 mg-1. Immunohistochemical studies localised these antigens in the hypodermis, the wall of the seminal receptacle and spermatozoa of adult worms. A lambda gt11 clone was isolated from an expression library of O. volvulus by immunoscreening. Sequence analysis revealed that it encoded a pi-class GST with 60% identity with Caenorhabditis elegans and up to 45% identity with mammalian pi-class GSTs. Antibodies affinity selected with recombinant GST demonstrated cross-reactivity between Litomosoides sigmodontis and O. volvulus GSTs.
Salinas
G
G
Department of Pathology, University of Cambridge, UK.
Braun
G
G
Taylor
D W
DW
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000032:analysis; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D003429:Cross Reactions / Q000276:immunology; D018509:DNA, Helminth / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics* / Q000302:isolation & purification*; D007124:Immunoenzyme Techniques; D008969:Molecular Sequence Data; D017181:Onchocerca volvulus / Q000201:enzymology* / Q000276:immunology; D016133:Polymerase Chain Reaction; D011817:Rabbits; D017386:Sequence Homology, Amino Acid
D000909:Antibodies, Helminth; D018509:DNA, Helminth; D005982:Glutathione Transferase