Rapid gating and anion permeability of an intracellular aquaporin.
Aquaporin (AQP) water-channel proteins are freely permeated by water but not by ions or charged solutes. Although mammalian aquaporins were believed to be located in plasma membranes, rat AQP6 is restricted to intracellular vesicles in renal epithelia. Here we show that AQP6 is functionally distinct from other known aquaporins. When expressed in Xenopus laevis oocytes, AQP6 exhibits low basal water permeability; however, when treated with the known water channel inhibitor, Hg2+, the water permeability of AQP6 oocytes rapidly rises up to tenfold and is accompanied by ion conductance. AQP6 colocalizes with H+-ATPase in intracellular vesicles of acid-secreting alpha-intercalated cells in renal collecting duct. At pH less than 5.5, anion conductance is rapidly and reversibly activated in AQP6 oocytes. Site-directed mutation of lysine to glutamate at position 72 in the cytoplasmic mouth of the pore changes the cation/anion selectivity, but leaves low pH activation intact. Our results demonstrate unusual biophysical properties of an aquaporin, and indicate that anion-channel function may now be explored in a protein with known structure.
Yasui
M
M
Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Hazama
A
A
Kwon
T H
TH
Nielsen
S
S
Guggino
W B
WB
Agre
P
P
D000818:Animals; D000838:Anions / Q000378:metabolism; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D002463:Cell Membrane Permeability; D006863:Hydrogen-Ion Concentration; D015640:Ion Channel Gating / Q000187:drug effects; D008564:Membrane Potentials; D008628:Mercury / Q000494:pharmacology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D012965:Sodium Chloride / Q000378:metabolism; D014867:Water / Q000378:metabolism; D014982:Xenopus laevis
D000838:Anions; C496025:Aqp2 protein, rat; C496043:Aqp6 protein, rat; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins; D011994:Recombinant Proteins; D014867:Water; D012965:Sodium Chloride; D008628:Mercury