Abstract
Permeabilities to glycerol and small non-electrolytes of three Aquaporin 1 CHIP (AQP1) water channels were measured in AQP1 cRNA-injected Xenopus laevis oocytes and in human AQP1 channels reconstituted in proteoliposomes. By an “osmotic” swelling assay, significant increases of ethylene glycol, glycerol and 1,3-propanediol apparent permeability coefficients (P′solutes) were found in oocytes expressing human, rat and frog AQP1. p-Chloromercuribenzene sulphonate (PCMBS) and CuSO4 inhibited, by 95% and 58% respectively, apparent glycerol permeability (P′ gly) in oocytes expressing human AQP1. pCMBS inhibition was reversed by β-mercaptoethanol and CuSO4 inhibition was partly reversed by the Cu2+-binding peptide Gly-Gly-His. Tritiated glycerol uptakes confirmed the augmented P′ gly value of AQP1 cRNA-injected oocytes. In contrast, no increases of urea, meso-erythritol, D- or L-threitol, xylitol and mannitol uptakes were detected. Stopped-flow light scattering experiments performed with human AQP1 proteoliposomes also revealed a much greater increase of P′ gly than did those with protein-free liposomes; the initial rate of proteoliposomes also swelling was inhibited by 96.2% with HgCl2 and by 72.5% with CuSO4. In AQP1 cRNA-injected oocytes and in proteoliposomes, the value of the glycerol reflection coefficient was 0.74–0.80, indicating that water and glycerol share the same pathway. All these results provide strong evidence that water and certain small solutes permeate the AQP1 channels expressed at the surface of X. laevis oocytes or reconstituted in proteoliposomes. The urea exclusion suggests that the selectivity of the AQP1 channels not only depends on the size of the solutes but probably also on their flexibility and their ability to form H-bonds.
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Abrami, L., Tacnet, F. & Ripoche, P. Evidence for a glycerol pathway through aquaporin 1 (CHIP28) channels. Pflügers Arch 430, 447–458 (1995). https://doi.org/10.1007/BF00373921
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DOI: https://doi.org/10.1007/BF00373921