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J. Biol. Chem., Vol. 265, Issue 26, 15375-15378, 09, 1990
RB Zhang, KA Logee and AS Verkman
The existence and identity of protein water transporters in biological
membranes has been uncertain. Osmotic water permeability (Pf) was measured
in defolliculated Xenopus oocytes microinjected with water or mRNA from
kidney cortex, kidney papilla, reticulocyte, brain, and muscle. Pf was
measured by quantitative image analysis from the time course of oocyte
swelling in response to an osmotic gradient. When assayed at 10 degrees C,
Pf in water-injected oocytes increased from (3.6 +/- 0.9) x 10(-4) cm/s
(S.D., n = 16) to 74 x 10(-4) cm/s with addition of amphotericin B, showing
absence of unstirred layers. At 48- 72 h after injection of 50 ng of
unfractionated mRNA, Pf (in cm/s x 10(- 4] was: 4.0 +/- 1.5 (rabbit brain,
n = 15), 4.2 +/- 1.8 (rabbit muscle, n = 10), 18.4 +/- 6.3 (rabbit
reticulocyte, n = 20), 16.1 +/- 5.6 (rat renal papilla, n = 24), 12.9 +/-
6.3 (rat renal cortex, n = 20), 14.4 +/- 6.1 (rabbit renal papilla, n =
15), and 11.8 +/- 3.4 (rabbit renal cortex, n = 8). In oocytes injected
with mRNA from rat renal papilla, Pf was inhibited reversibly by 0.3 mM
HgCl2 (4.1 +/- 1.6, n = 10); expressed water channels from kidney and red
cell had activation energies of less than 4 kcal/mol. These results show
functional oocyte expression of water channels from red cell, kidney
proximal tubule (cortex), and the vasopressin-sensitive kidney collecting
tubule (papilla), indicating that water channels are proteins, and
providing an approach for the expression cloning of water channels.
Expression of mRNA coding for kidney and red cell water channels in Xenopus oocytes
Department of Medicine, University of California, San Francisco 94143- 0532.
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