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Quantification of the Intracellular Life Time of Water Molecules to Measure Transport Rates of Human Aquaglyceroporins

Journal article
Authors Madelene Palmgren
Malin Hernebring
S. Eriksson
K. Elbing
Cecilia Geijer
S. Lasic
Peter Dahl
J. S. Hansen
D. Topgaard
Karin Lindkvist-Petersson
Published in Journal of Membrane Biology
Volume 250
Issue 6
Pages 629-639
ISSN 0022-2631
Publication year 2017
Published at Department of Chemistry and Molecular Biology
Pages 629-639
Language en
Links dx.doi.org/10.1007/s00232-017-9988-...
Keywords Aquaporin, Aquaglyceroporin, Water transport, NMR, P. pastoris, pichia-pastoris, glycerol, aquaporins, expression, metabolism, system, aqp9, Biochemistry & Molecular Biology, Cell Biology, Physiology
Subject categories Cell biology

Abstract

Orthodox aquaporins are transmembrane channel proteins that facilitate rapid diffusion of water, while aquaglyceroporins facilitate the diffusion of small uncharged molecules such as glycerol and arsenic trioxide. Aquaglyceroporins play important roles in human physiology, in particular for glycerol metabolism and arsenic detoxification. We have developed a unique system applying the strain of the yeast Pichia pastoris, where the endogenous aquaporins/aquaglyceroporins have been removed and human aquaglyceroporins AQP3, AQP7, and AQP9 are recombinantly expressed enabling comparative permeability measurements between the expressed proteins. Using a newly established Nuclear Magnetic Resonance approach based on measurement of the intracellular life time of water, we propose that human aquaglyceroporins are poor facilitators of water and that the water transport efficiency is similar to that of passive diffusion across native cell membranes. This is distinctly different from glycerol and arsenic trioxide, where high glycerol transport efficiency was recorded.

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