Functional analysis of novel aquaporins from Fasciola gigantica

Mol Biochem Parasitol. 2011 Feb;175(2):144-53. doi: 10.1016/j.molbiopara.2010.10.010. Epub 2010 Nov 10.

Abstract

Fascioliasis, caused by liver flukes of the genus Fasciola, is an important disease of ruminants. In order to identify a potential new drug target we have studied aquaporin (AQP) in Fasciola gigantica. AQPs facilitate the transport of water, glycerol and other small solutes across biological membranes. The structure, function, and pathology of AQPs have been extensively studied in mammals but data for AQPs from trematodes is still limited. In the present study, we have functionally characterized two closely related AQP isoforms, FgAQP-1 and FgAQP-2, from the trematode F. gigantica. Immunohistochemical analysis located the FgAQPs in the tegumental cells, their processes and the tegument itself. In addition, they were present in the epithelial linings of testes and ovary. Expression in Xenopus oocytes of these FgAQPs increased osmotic water permeability 3-4-fold but failed to increase glycerol and urea permeability. AQPs have two highly conserved NPA motifs that are important for the function of the channel pore. In FgAQP-1 and FgAQP-2 the first NPA motif is changed to TAA. Substitution of Thr with Asn in the TAA motif of FgAQP-1 increased its water permeability twofold but did not affect urea and glycerol impermeability while the substitution at the pore mouth of Cys204 by Tyr caused loss of water permeability. In addition, the FgAQPs did not increase methylamine and ammonia permeability after expression in yeast. In comparison to rat AQP-1 the described FgAQPs showed low water permeability and further in vivo analyses are necessary to determine their contribution to osmoregulation in Fasciola.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Amino Acid Substitution / genetics
  • Ammonia / metabolism
  • Animals
  • Aquaporins / genetics
  • Aquaporins / metabolism*
  • Biological Transport
  • Conserved Sequence
  • DNA, Helminth / chemistry
  • DNA, Helminth / genetics
  • Fasciola / chemistry
  • Fasciola / enzymology*
  • Fasciola / genetics
  • Glycerol / metabolism
  • Immunohistochemistry
  • Methylamines / metabolism
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Oocytes / parasitology
  • Phylogeny
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Urea / metabolism
  • Water / metabolism
  • Xenopus / parasitology

Substances

  • Aquaporins
  • DNA, Helminth
  • Methylamines
  • Protein Isoforms
  • Water
  • Ammonia
  • Urea
  • methylamine
  • Glycerol

Associated data

  • GENBANK/HM748644
  • GENBANK/HM748645