Cloning of a novel water and urea-permeable aquaporin from mouse expressed strongly in colon, placenta, liver, and heart.
An aquaporin-type water channel was isolated from mouse based on homology to known aquaporins. A 1447 bp cDNA was sequenced (designated AQP8) with a 783 bp open reading frame encoding a 261 amino acid hydrophobic protein which contained the conserved NPA motifs of MIP family members. Amino acid alignment showed greatest homology of AQP8 to plant water channel gamma TIP (38% identity) followed by mammalian water channels AQP4 (32%) and AQP2 (31%). Northern blot analysis indicated a 1.7 kb transcript expressed strongly in placenta > colon > liver approximately heart. RT-PCR with AQP8-specific primers and Southern blot analysis showed AQP8 transcript in the above tissues and in pancreas, lung, kidney, submandibular gland, diaphragm, testis, spleen, stomach and brain. Expression of AQP8 cRNA in Xenopus oocytes increased osmotic water permeability from (0.8 +/- 0.1) x 10(-3) cm/s to (22 +/- 3) x 10(-3) cm/s (10 degrees C) in a mercurial-sensitive manner. AQP8 was also permeable to urea but not to glycerol. Normalization for oocyte plasma membrane expression using cMyc-tagged AQP8 indicated a single channel water permeability of 8.2 x 10(-14) cm3/s. AQP8 is unique among the water channels in terms of its urea permeability and its strong expression in gastrointestinal organs, placenta and heart.
Ma
T
T
Department of Medicine, University of California, San Francisco 94143-0521, USA.
Yang
B
B
Verkman
A S
AS
D000595:Amino Acid Sequence; D000818:Animals; D020346:Aquaporins; D001483:Base Sequence; D003001:Cloning, Molecular; D003106:Colon / Q000378:metabolism*; D005260:Female; D007473:Ion Channels / Q000096:biosynthesis / Q000737:chemistry / Q000502:physiology*; D008099:Liver / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism; D009865:Oocytes / Q000502:physiology; D010920:Placenta / Q000378:metabolism*; D011247:Pregnancy; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D014508:Urea / Q000378:metabolism*; D014982:Xenopus laevis
D020346:Aquaporins; D007473:Ion Channels; D012333:RNA, Messenger; D011994:Recombinant Proteins; C108330:aquaporin 8; D014508:Urea