Evidence of transferrin binding sites on the surface of Leishmania promastigotes.
A glycoprotein of 78,000 molecular mass (78 kDa), associated with the membrane of Leishmania infantum promastigotes, was identified and immunopurified by monoclonal antibody (mAb) LD9 produced against isolated membrane preparations. mAb LD9 was subsequently found to bind to human transferrin, also of 78 kDa. Binding of LD9 to transferrin was completely abolished when the mAb was preabsorbed by Leishmania membranes, thereby indicating that the 78-kDa Leishmania membrane-associated glycoprotein and transferrin have common antigenic epitope(s). The 78-kDa Leishmania membrane-associated protein was released in soluble nonaggregated form by mild treatment with acetic acid saline. Anti-transferrin polyclonal antibodies, recognized both the membrane-associated and the soluble form of the 78-kDa glycoprotein. The 78-kDa soluble form was characterized further as an iron-containing protein. The above data combined with iron uptake by promastigotes as demonstrated by the Prussian blue reaction indicate that the 78-kDa Leishmania membrane-associated glycoprotein is transferrin. The binding of 125I-human transferrin to Leishmania-purified membrane preparations was then investigated. The results indicate the presence of a high affinity saturable binding site (Kd = 2.2 10(-8) M) that is specific for transferrin. We suggest that the 78-kDa glycoprotein recognized by mAb LD9 is transferrin that binds to the surface of Leishmania promastigotes via a transferrin receptor.
Laboratory of Biochemistry, Hellenic Pasteur Institute, Athens, Greece.
C S
CS
Voyiatzaki
K P
KP
Soteriadou
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D002846:Chromatography, Affinity; D000939:Epitopes / Q000032:analysis; D007700:Kinetics; D007893:Leishmania donovani / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D011990:Receptors, Transferrin / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D014168:Transferrin / Q000378:metabolism*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes; D008562:Membrane Glycoproteins; D011990:Receptors, Transferrin; D014168:Transferrin