Biochemical characterization of Leishmania major aquaglyceroporin LmAQP1: possible role in volume regulation and osmotaxis.
The Leishmania major aquaglyceroporin, LmAQP1, is responsible for the transport of trivalent metalloids, arsenite and antimonite. We have earlier shown that downregulation of LmAQP1 provides resistance to trivalent antimony compounds whereas increased expression of LmAQP1 in drug-resistant parasites can reverse the resistance. In this paper we describe the biochemical characterization of LmAQP1. Expression of LmAQP1 in Xenopus oocytes rendered them permeable to water, glycerol, methylglyoxal, dihydroxyacetone and sugar alcohols. The transport property of LmAQP1 was severely affected when a critical Arg230, located inside the pore of the channel, was altered to either alanine or lysine. Immunofluorescence and immuno-electron microscopy revealed LmAQP1 to be localized to the flagellum of Leishmania promastigotes and in the flagellar pocket membrane and contractile vacuole/spongiome complex of amastigotes. This is the first report of an aquaglyceroporin being localized to the flagellum of any microbe. Leishmania promastigotes and amastigotes expressing LmAQP1 could regulate their volume in response to hypoosmotic stress. Additionally, Leishmania promastigotes overexpressing LmAQP1 were found to migrate faster towards an osmotic gradient. These results taken together suggest that Leishmania LmAQP1 has multiple physiological roles, being involved in solute transport, volume regulation and osmotaxis.
Figarella
Katherine
K
Department of Biochemistry and Molecular Biology, Wayne State University, School of Medicine, Detroit, MI, USA.
Uzcategui
Nestor L
NL
Zhou
Yao
Y
LeFurgey
Ann
A
Ouellette
Marc
M
Bhattacharjee
Hiranmoy
H
Mukhopadhyay
Rita
R
D000818:Animals; D051397:Aquaglyceroporins / Q000378:metabolism*; D001120:Arginine / Q000235:genetics; D001692:Biological Transport; D002463:Cell Membrane Permeability; D048429:Cell Size; D002633:Chemotaxis; D007893:Leishmania donovani / Q000166:cytology / Q000254:growth & development / Q000648:ultrastructure; D018320:Leishmania major / Q000378:metabolism*; D008018:Life Cycle Stages; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D009865:Oocytes; D009995:Osmosis; D015800:Protozoan Proteins / Q000378:metabolism*; D014981:Xenopus
D051397:Aquaglyceroporins; D050505:Mutant Proteins; D015800:Protozoan Proteins; D001120:Arginine