Biochemical and immunochemical characterization of a P-type ATPase from Leishmania donovani promastigote plasma membrane.
An ATPase on the plasma membrane of Leishmania donovani has been characterized. An antiserum, generated against ATPase active bands from native gels, was specific for a 105 kDa protein in promastigotes. However, in plasma membrane preparations a 70 kDa protein is also recognized, suggesting proteolysis of the intact 105 kDa protein or the presence of a second similar ATPase. [gamma-32P]ATP phosphorylates two proteins (105 kDa and 70 kDa) in promastigotes and plasma membranes. Both proteins form a transient phosphorylated intermediate, characteristic of a P-type ATPase. Immunostaining of permeabilized parasites shows diffuse staining of the surface of promastigotes and amastigotes, which is consistent with a plasma membrane protein. The antiserum immunoprecipitates a 70 kDa [14C]DCCD binding protein from whole cells and plasma membranes of promastigotes. Furthermore, the antiserum immunoprecipitates a 105 kDa and 70 kDa protein which can be subsequently phosphorylated. These results indicate the presence of a 105 kDa P-type ATPase on the L. donovani plasma membrane which is similar to the mammalian and fungal cation pumps.
Anderson
S A
SA
Department of Biological Sciences, University of Cincinnati, OH 45221.
Mukkada
A J
AJ
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D002458:Cell Fractionation; D002462:Cell Membrane / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D016650:Fluorescein-5-isothiocyanate; D005455:Fluorescent Antibody Technique; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting; D007893:Leishmania donovani / Q000201:enzymology*; D008274:Magnesium; D008345:Manganese; D011233:Precipitin Tests
D007106:Immune Sera; D008345:Manganese; D000251:Adenosine Triphosphatases; D016650:Fluorescein-5-isothiocyanate; D008274:Magnesium