title: Characterization of plasma membrane bound inorganic pyrophosphatase from Leishmania donovani promastigotes and amastigotes.
abstract: BACKGROUND
  Currently, a major problem in the management of visceral leishmaniasis or kala-azar, especially in the Indian subcontinent, is the growing unresponsiveness to conventional antimonial therapy. Membrane bound pyrophophatase (PPases) do not exist in plasma membrane from mammals. Thus, H(+)-PPases from Leishmania plasma membrane might be potential target in rational chemotherapy of the disease caused by Leishmania parasites.
  
  
  OBJECTIVE
  To characterize the activities of inorganic pyrophophatase (PPase) in the plasma membrane of Leishmania donovani promastigote and amastigote.
  
  
  METHODS
  Culture method of promastigote and amastigote were developed. We assayed PPase activity in isolated plasma membrane of L. donovani.
  
  
  RESULTS
  We characterized K(+)-PPase present in the plasma membrane of Leishmania donovani and investigated its possible role in the survival of promastigote and amastigote. PPase activity was stimulated by K(+) ions and sodium orthovanadate, inhibited by pyrophosphate analogs imidodiphosphate and alendronate, KF, DCCD, thiol reagent parachloromercuribenzenesulfonate (PCMBS), N-ethylmaliemide (NEM), phenylarsineoxide, ABC superfamily transport modulator verapamil and was also by F(1)F(o)-ATPase inhibitor quercetin.
  
  
  CONCLUSIONS
  We conclude that there are significant differences within promastigote, amastigote and mammalian host in cytosolic pH homeostasis to merit the inclusion of PPase transporter as putative targets for rational drug design.
authors.affiliation: Division of Medicinal Biochemistry, Department of Pharmaceutical Technology, India.
authors.forename: S S
authors.initials: SS
authors.lastname: Sen
authors.forename: N R
authors.initials: NR
authors.lastname: Bhuyan
authors.forename: Tanmoy
authors.initials: T
authors.lastname: Bera
mesh_terms: D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology*; D015195:Drug Design; D004352:Drug Resistance, Microbial; D043564:Inorganic Pyrophosphatase / Q000187:drug effects / Q000378:metabolism*; D007893:Leishmania donovani / Q000378:metabolism*; D007896:Leishmaniasis / Q000188:drug therapy
chemical_list: D000981:Antiprotozoal Agents; D043564:Inorganic Pyrophosphatase