title: Biochemical and immunochemical characterization of a P-type ATPase from Leishmania donovani promastigote plasma membrane.
abstract: 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.
authors.affiliation: Department of Biological Sciences, University of Cincinnati, OH 45221.
authors.forename: S A
authors.initials: SA
authors.lastname: Anderson
authors.forename: A J
authors.initials: AJ
authors.lastname: Mukkada
mesh_terms: 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
chemical_list: D007106:Immune Sera; D008345:Manganese; D000251:Adenosine Triphosphatases; D016650:Fluorescein-5-isothiocyanate; D008274:Magnesium