Chemical crosslinking of Plasmodium falciparum glycoprotein, Pf200 (190-205 kDa), to the S-antigen at the merozoite surface.
Merozoites were isolated from Plasmodium falciparum cultures labeled with [3H]mannose and [35S]methionine and treated with a cleavable homobifunctional crosslinker, dithiobis(succinimidyl) propionate. The crosslinked complexes were immunoprecipitated with Mab.5B1 directed against the major merozoite surface glycoprotein. Pf200 (MW 190-205), and reduced with dithiothreitol. Crosslinked immunocomplexes did not contain the second major merozoite surface glycoprotein, Pf50 (MW 45-55 kDa), or other major [35S]methionine-labeled proteins, except for a weakly labeled protein of 150 kDa. Crosslinked complexes immunoprecipitated with Mab.5B1 and then reduced with DTT were immunoblotted with antibody directed against three soluble P. falciparum antigens, a serine-rich antigen known as Pf126 or SERA, the S-antigen, and GBP-130. The 150-kDa S-antigen was readily detected in crosslinked immunocomplexes with Pf200. The SERA antigen, although crosslinked under these conditions, was not detected in association with Pf200 nor was GBP-130.
Perkins
M E
ME
Laboratory of Biochemical Parasitology, Rockefeller University, New York, New York 10021.
Rocco
L J
LJ
D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology; D003432:Cross-Linking Reagents; D006023:Glycoproteins / Q000276:immunology*; D015151:Immunoblotting; D008565:Membrane Proteins / Q000276:immunology; D010963:Plasmodium falciparum / Q000276:immunology*; D011233:Precipitin Tests; D015800:Protozoan Proteins / Q000276:immunology*
D000953:Antigens, Protozoan; D000954:Antigens, Surface; D003432:Cross-Linking Reagents; D006023:Glycoproteins; D008565:Membrane Proteins; D015800:Protozoan Proteins; C047712:S-antigen, Plasmodium falciparum