Plasmodium falciparum: identification and localization of a knob protein antigen expressed by a cDNA clone.
Differential screening of cDNA libraries constructed from knobby and predominantly knobless Plasmodium falciparum isolates, identified the sequence SD17. Chromosome blotting experiments have shown that this sequence, which is located on chromosome 2 of most isolates, was deleted in the cloned parasite line E12 of the FCQ27/PNG isolate. Here we show that erythrocytes infected with the SD17-containing cloned line D10 have typical knob structures on their surfaces, whereas those infected with the line E12 lack knobs. An expression clone was constructed from SD17 and used to affinity purify antibodies from the sera of individuals living in areas of Papua New Guinea where malaria is endemic. The antibodies reacted in immunoblotting experiments with a single polypeptide that varied in Mr from 85,000 to 105,000 among different isolates. The antigen was not expressed in the knobless clone E12. Postembedding immunoelectron microscopy showed localization of the antigen over the knobs of FC27 and two other isolates, largely on the cytoplasmic side. We conclude that the parasite antigen corresponding to clone SD17 is a knob protein.
D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004910:Erythrocyte Membrane / Q000032:analysis / Q000648:ultrastructure; D004912:Erythrocytes / Q000469:parasitology / Q000648:ultrastructure; D006801:Humans; D007158:Immunologic Techniques; D008854:Microscopy, Electron; D010455:Peptides / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D010963:Plasmodium falciparum / Q000032:analysis / Q000235:genetics* / Q000276:immunology / Q000648:ultrastructure; D015800:Protozoan Proteins
D000953:Antigens, Protozoan; D010455:Peptides; D015800:Protozoan Proteins; C042968:knob protein, Plasmodium falciparum; D004247:DNA