title: Expression and functional characterization of a Plasmodium falciparum Ca2+-ATPase (PfATP4) belonging to a subclass unique to apicomplexan organisms.
abstract: We have obtained a full-length P type ATPase sequence (PfATP4) encoded by Plasmodium falciparum and expressed PfATP4 in Xenopus laevis oocytes to study its function. Comparison of the hitherto incomplete open reading frame with other Ca(2+)-ATPase sequences reveals that PfATP4 differs significantly from previously defined categories. The Ca(2+)-dependent ATPase activity of PfATP4 is stimulated by a much broader range of [Ca(2+)](free) (3.2-320 micrometer) than are an avian SERCA1 pump or rabbit SERCA 1a (maximal activity < 10 micrometer). The activity of PfATP4 is resistant to inhibition by ouabain (200 micrometer) or thapsigargin (0.8 micrometer) but is inhibited by vanadate (1 mM) or cyclopiazonic acid (1 microM). We used a quantitative polymerase chain reaction to assay expression of mRNA encoding PfATP4 relative to that for beta-tubulin in synchronized asexual stages and found variable expression throughout the life cycle with a maximal 5-fold increase in meronts compared with ring stages. This analysis suggests that PfATP4 defines a novel subclass of Ca(2+)-ATPases unique to apicomplexan organisms and therefore offers potential as a drug target.
authors.affiliation: Department of Infectious Diseases, St. George's Hospital Medical School, Cranmer Terrace, London SW17 ORE, United Kingdom. s.krishna@sghms.ac.uk
authors.forename: S
authors.initials: S
authors.lastname: Krishna
authors.forename: C
authors.initials: C
authors.lastname: Woodrow
authors.forename: R
authors.initials: R
authors.lastname: Webb
authors.forename: J
authors.initials: J
authors.lastname: Penny
authors.forename: K
authors.initials: K
authors.lastname: Takeyasu
authors.forename: M
authors.initials: M
authors.lastname: Kimura
authors.forename: J M
authors.initials: JM
authors.lastname: East
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000252:Calcium-Transporting ATPases / Q000032:analysis* / Q000096:biosynthesis* / Q000145:classification / Q000235:genetics; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment; D017421:Sequence Analysis
chemical_list: D000252:Calcium-Transporting ATPases