title: The Nramp orthologue of Cryptococcus neoformans is a pH-dependent transporter of manganese, iron, cobalt and nickel.
abstract: Cryptococcus neoformans is an important human opportunistic pathogen and a facultative intracellular parasite, particularly in HIV-infected individuals. Little is known about metal ion transport in this organism. C. neoformans encodes a single member of the Nramp (natural resistance-associated macrophage protein) family of bivalent cation transporters, known as Cramp, which we have cloned and expressed in Xenopus laevis oocytes and Spodoptera frugiperda Sf 21 insect cells. Cramp induces saturable transport of a broad range of bivalent transition series cations, including Mn2+, Fe2+, Co2+ and Ni2+. Maximal cation transport occurs at pH 5.5-6.0, consistent with the proton gradient-based energetics of other Nramp orthologues. Mn2+ transport is diminished in the presence of 140 mM Na+, compatible with a Na+ slippage mechanism proposed for the Saccharomyces cerevisiae Nramp orthologue Smf1p. Cramp resembles Smf1p with respect to predicted membrane topology, substrate specificity and pH dependence, but differs in terms of its apparent affinity for Mn2+ and negligible inhibition by Zn2+. Cramp is the first Nramp orthologue from a fungal pathogen to be functionally characterized. Insights afforded by these findings will allow the formulation of new hypotheses regarding the role of metal ions in the pathophysiology of cryptococcosis.
authors.affiliation: Department of Cellular and Molecular Medicine (Infectious Diseases), St George's Hospital Medical School, Cranmer Terrace, London SW17 ORE, UK.
authors.forename: Daniel
authors.initials: D
authors.lastname: Agranoff
authors.forename: Lauren
authors.initials: L
authors.lastname: Collins
authors.forename: David
authors.initials: D
authors.lastname: Kehres
authors.forename: Tom
authors.initials: T
authors.lastname: Harrison
authors.forename: Michael
authors.initials: M
authors.lastname: Maguire
authors.forename: Sanjeev
authors.initials: S
authors.lastname: Krishna
mesh_terms: D000818:Animals; D027682:Cation Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002413:Cations, Divalent / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002712:Chlorides / Q000494:pharmacology; D003035:Cobalt / Q000378:metabolism*; D003455:Cryptococcus neoformans / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D017136:Ion Transport; D007501:Iron / Q000378:metabolism*; D007700:Kinetics; D008345:Manganese / Q000378:metabolism*; D009532:Nickel / Q000378:metabolism*; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D017422:Sequence Analysis, DNA; D012964:Sodium / Q000494:pharmacology; D018411:Spodoptera; D013379:Substrate Specificity; D014981:Xenopus; D000099173:Natural Resistance-associated Macrophage Protein 1
chemical_list: D027682:Cation Transport Proteins; D002413:Cations, Divalent; D002712:Chlorides; D003035:Cobalt; D007501:Iron; D008345:Manganese; D009532:Nickel; D012964:Sodium; D000099173:Natural Resistance-associated Macrophage Protein 1