title: Expression, purification and biochemical characterization of recombinant Ca-dependent protein kinase 2 of the malaria parasite Plasmodium falciparum.
abstract: Calcium-dependent protein kinases (CDPKs) are serine/threonine kinases that react in response to calcium which functions as a trigger for several mechanisms in plants and invertebrates, but not in mammals. Recent structural studies have defined the role of calcium in the activation of CDPKs and have elucidated the important structural changes caused by calcium in order to allow the kinase domain of CDPK to bind and phosphorylate the substrate. However, the role of autophosphorylation in CDPKs is still not fully understood. In Plasmodium falciparum, seven CDPKs have been identified by sequence comparison, and four of them have been characterized and assigned to play a role in parasite motility, gametogenesis and egress from red blood cells. Although PfCDPK2 was already discovered in 1997, little is known about this enzyme and its metabolic role. In this work, we have expressed and purified PfCDPK2 at high purity in its unphosphorylated form and characterized its biochemical properties. Moreover, propositions about putative substrates in P. falciparum are made based on the analysis of the phosphorylation sites on the artificial substrate myelin basic protein (MBP).
authors.affiliation: Pharmaceutical Biochemistry Group, School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Geneva, Switzerland.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Lauciello
authors.forename: Barbara
authors.initials: B
authors.lastname: Kappes
authors.forename: Leonardo
authors.initials: L
authors.lastname: Scapozza
authors.forename: Remo
authors.initials: R
authors.lastname: Perozzo
mesh_terms: D000818:Animals; D010766:Phosphorylation; D010963:Plasmodium falciparum / Q000201:enzymology*; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D017871:Calcium-Calmodulin-Dependent Protein Kinases
chemical_list: D011494:Protein Kinases; D015800:Protozoan Proteins; D017871:Calcium-Calmodulin-Dependent Protein Kinases
keywords: CDPK2; Enzyme kinetics; Plasmodium falciparum; Protein kinase