The ring-infected erythrocyte surface antigen protein of Plasmodium falciparum is phosphorylated upon association with the host cell membrane.
The ring-infected erythrocyte surface antigen (RESA) is a 155-kDa malarial polypeptide which is released from merozoites and becomes associated with the erythrocyte membrane at the time of invasion. Inside-out vesicles (IOVs) prepared from Plasmodium falciparum-infected erythrocytes contain RESA, presumably bound to the membrane skeleton, as it is largely insoluble in Triton X-100. When these IOVs were incubated with [gamma-32P]ATP, a 155-kDa polypeptide was labeled in IOVs from infected, but not from uninfected erythrocytes. Immunoprecipitation using specific rabbit antisera confirmed that RESA is indeed a phosphoprotein. Phosphoamino acid analysis revealed phosphoserine and a small amount of phosphothreonine, but no phosphotyrosine. Labeling of intact parasitized erythrocytes with inorganic [32P]phosphate for several hours in culture resulted in RESA in Triton-insoluble extracts being phosphorylated. Labeling of synchronized parasites showed that RESA was phosphorylated only when it became associated with the erythrocyte membrane, and although RESA was abundant in mature parasites, it was not phosphorylated. RESA, released into the culture supernatants during the growth of P. falciparum, bound to IOVs prepared from normal uninfected erythrocytes, and subsequent labeling with [gamma-32P]ATP resulted in the phosphorylation of RESA. The evidence suggests that RESA is phosphorylated by an erythrocyte membrane kinase and probably not by a parasite-encoded enzyme.
Foley
M
M
Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
Murray
L J
LJ
Anders
R F
RF
D000255:Adenosine Triphosphate / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000953:Antigens, Protozoan / Q000378:metabolism*; D000954:Antigens, Surface / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000378:metabolism* / Q000469:parasitology; D015151:Immunoblotting; D010761:Phosphorus Radioisotopes / Q000378:metabolism; D010766:Phosphorylation; D010963:Plasmodium falciparum / Q000502:physiology*; D011233:Precipitin Tests; D015800:Protozoan Proteins; D012694:Serine / Q000378:metabolism; D013912:Threonine / Q000378:metabolism
D000596:Amino Acids; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D010761:Phosphorus Radioisotopes; D015800:Protozoan Proteins; C050256:ring-infected erythrocyte surface antigen (RESA), Plasmodium falciparum; D013912:Threonine; D012694:Serine; D000255:Adenosine Triphosphate