Biochemical properties of a 24 kilodalton membrane glycoprotein antigen complex from Schistosoma mansoni.
Antibodies were affinity purified from crude antiserum by elution from the 24 kDa region of preparative one-dimensional Western blots containing immobilized adult Schistosoma mansoni inner bilayer membrane proteins. They were shown to be specific for a single acidic polypeptide complex, Smgp24, following immunoblotting from two-dimensional polyacrylamide gels. These antibodies were then used to detect the presence of the Smgp24 complex in fractions prepared from lectin affinity chromatography, phase separation in Triton X-114 and chemical and enzymatic carbohydrate modification treatments. The 24 kDa antigen was bound and specifically eluted from both concanavalin A and lentil lectin affinity matrices. In addition, the electrophoretic mobility of the antigen was shifted to approximately 20 kDa after treatment with endoglycosidase F and N-glycanase, but was not appreciably altered following treatment with endoglycosidase H, neuraminidase, or sodium meta-periodate. The 20 kDa species produced by endoglycosidase F or N-glycanase treatment no longer bound to the lectin affinity resins. The Smgp24 complex also partitioned almost quantitatively into the detergent-enriched phase after phase separation in Triton X-114 solutions. These results indicate that the Smgp24 complex is an antigenic integral membrane glycoprotein and may consist of a single polypeptide backbone which is extensively post- or co-translationally modified.
Karcz
S R
SR
Department of Zoology, University of Western Ontario, London, Canada.
Barnard
B J
BJ
Podesta
R B
RB
D000818:Animals; D000947:Antigens, Helminth / Q000032:analysis* / Q000378:metabolism; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D037102:Lectins / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000378:metabolism; D012550:Schistosoma mansoni / Q000276:immunology*
D000947:Antigens, Helminth; D037102:Lectins; D008562:Membrane Glycoproteins