Purified E255L Mutant SERCA1a and Purified PfATP6 Are Sensitive to SERCA-type Inhibitors but Insensitive to Artemisinins
Autor: | Marc le Maire, Bertrand Arnou, Alexandre Pozza, Christine Jaxel, Delphine Cardi, Estelle Marchal, Jesper V. Møller, Sanjeev Krishna, Johannes D. Clausen, Jens Peter Andersen |
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Přispěvatelé: | Système membranaires, photobiologie, stress et détoxication (SMPSD), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS) |
Předmět: |
Artemisinins
Thapsigargin SERCA 030231 tropical medicine Mutant ATPases Mutation Missense Calcium-Transporting ATPases Saccharomyces cerevisiae Biochemistry Sarcoplasmic Reticulum Calcium-Transporting ATPases Antimalarials 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Membrane Biology Chlorocebus aethiops parasitic diseases medicine Animals [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Enzyme Inhibitors Molecular Biology 030304 developmental biology 0303 health sciences COS cells biology Drug Action Membrane Proteins Plasmodium falciparum Cell Biology biology.organism_classification Molecular biology 3. Good health Mechanism of action chemistry COS Cells Calcium ATPase Mutant Proteins Rabbits medicine.symptom Cyclopiazonic acid |
Zdroj: | Aarhus University Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2010, pp.26406-26416 Cardi, D, Pozza, A, Arnou, B, Marchal, E L, Clausen, J D, Andersen, J P, Krishna, S, Møller, J V, le Maire, M & Jaxel, C 2010, ' Purified E255L Mutant SERCA1a and Purified PfATP6 Are Sensitive to SERCA-type Inhibitors but Insensitive to Artemisinins. ', Journal of Biological Chemistry, vol. 285, no. 34, pp. 26406-26416 . The Journal of Biological Chemistry Journal of Biological Chemistry, 2010, 285 (34), pp.26406-26416. ⟨10.1074/jbc.M109.090340⟩ |
ISSN: | 0021-9258 1083-351X |
Popis: | The antimalarial drugs artemisinins have been described as inhibiting Ca(2+)-ATPase activity of PfATP6 (Plasmodium falciparum ATP6) after expression in Xenopus oocytes. Mutation of an amino acid residue in mammalian SERCA1 (Glu(255)) to the equivalent one predicted in PfATP6 (Leu) was reported to induce sensitivity to artemisinin in the oocyte system. However, in the present experiments, we found that artemisinin did not inhibit mammalian SERCA1a E255L either when expressed in COS cells or after purification of the mutant expressed in Saccharomyces cerevisiae. Moreover, we found that PfATP6 after expression and purification from S. cerevisiae was insensitive to artemisinin and significantly less sensitive to thapsigargin and 2,5-di(tert-butyl)-1,4-benzohydroquinone than rabbit SERCA1 but retained higher sensitivity to cyclopiazonic acid, another type of SERCA1 inhibitor. Although mammalian SERCA and purified PfATP6 appear to have different pharmacological profiles, their insensitivity to artemisinins suggests that the mechanism of action of this class of drugs on the calcium metabolism in the intact cell is complex and cannot be ascribed to direct inhibition of PfATP6. Furthermore, the successful purification of PfATP6 affords the opportunity to develop new antimalarials by screening for inhibitors against PfATP6. |
Databáze: | OpenAIRE |
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