Relative protective properties of three membrane glycoprotein fractions from Haemonchus contortus.
Jacalin lectin was used as a ligand to isolate a fraction containing two distinct protective antigens from detergent extracts of membranes from Haemonchus contortus. The first antigen was identified as a complex which appeared very similar to Haemonchus galactose-containing glycoprotein (H-gal-GP), which is a previously described protective protease complex, except that it was substantially depleted of one of the main H-gal-GP components, a 230 kDa metallopeptidase-containing band. The new complex was termed Haemonchus sialylated galactose-containing glycoprotein (H-sialgal-GP), because it bound to jacalin but not to peanut lectin and only jacalin will bind the sialylated form of galactosyl (beta-1, 3) N-acetylgalactosamine. Two protection trials with sheep showed that H-sialgal-GP and H-gal-GP were equally efficacious, reducing numbers of Haemonchus eggs by between 86% and 93% and worms by between 52% and 75%, respectively. The second jacalin-binding protective antigen fraction was separated from H-sialgal-GP by ion exchange and gel filtration chromatography. It was greatly enriched for two proteins termed p46 and p52 according to their apparent molecular weights. Immunization of sheep with these proteins gave protection values of 78% for eggs and 33% for worms, which are significantly lower than those obtained with either H-gal-GP or H-sialgal-GP. N-terminal amino acid sequence data from p46 and p52 showed that both proteins were closely related to a previously described 45 kDa Haemonchus membrane protein, which had conferred protection against Haemonchus in guinea-pigs.
Smith
W D
WD
Moredun Research Institute, International Research Centre, Pentland Science Park, Bush Loan, Penicuik EH26 OPZ, UK.
Smith
S K
SK
Pettit
D
D
Newlands
G F
GF
Skuce
P J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006189:Haemonchus / Q000737:chemistry / Q000276:immunology*; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007114:Immunization; D007422:Intestines / Q000737:chemistry; D037102:Lectins; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010270:Parasite Egg Count; D037121:Plant Lectins; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012756:Sheep / Q000276:immunology / Q000469:parasitology*; D013997:Time Factors
D000947:Antigens, Helminth; D015801:Helminth Proteins; D037102:Lectins; D008562:Membrane Glycoproteins; D037121:Plant Lectins; C043696:jacalin; D010450:Endopeptidases; C406387:H-sialgal-GP protein, Haemonchus contortus; C119206:galactose-containing glycoprotein, Haemonchus contortus