Characterization of the cross-reacting determinant (CRD) of the glycosyl-phosphatidylinositol membrane anchor of Trypanosoma brucei variant surface glycoprotein.
The cross-reacting determinant (CRD epitope) of the glycosyl-phosphatidylinositol (GPI) membrane anchor of Trypanosoma brucei variant surface glycoprotein has been analysed by selective chemical and enzymic modification of the isolated GPI structure combined with the use of a competitive ELISA inhibition assay for the detection of CRD epitopes. The data show that the CRD consists of at least three overlapping epitopes involving different regions of the molecule including the inositol 1,2-cyclic phosphate, the non-N-acetylated-glucosamine residue and the galactose branch. Although the presence of all three of these structural features is required for quantitative binding of anti-CRD antibodies in ELISA and Western blotting, the Western blot reaction obtained in the presence of any one epitope is still significant. The use of anti-CRD antibodies for the detection of GPI anchors is discussed.
Zamze
S E
SE
Department of Biochemistry, University of Oxford, England.
Ferguson
M A
MA
Collins
R
R
Dwek
R A
RA
Rademacher
T W
TW
D000818:Animals; D000906:Antibodies / Q000032:analysis; D001665:Binding Sites; D001667:Binding, Competitive; D015153:Blotting, Western; D002462:Cell Membrane / Q000032:analysis; D002999:Clone Cells / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D006017:Glycolipids / Q000032:analysis* / Q000276:immunology; D017261:Glycosylphosphatidylinositols; D008563:Membrane Lipids / Q000032:analysis*; D010716:Phosphatidylinositols / Q000032:analysis* / Q000276:immunology; D013329:Structure-Activity Relationship; D014346:Trypanosoma brucei brucei / Q000032:analysis*; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000032:analysis*
D000906:Antibodies; D000939:Epitopes; D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D008563:Membrane Lipids; D010716:Phosphatidylinositols; D014643:Variant Surface Glycoproteins, Trypanosoma