Reversible, temperature-dependent supramolecular assembly of aquaporin-4 orthogonal arrays in live cell membranes.

Biophysical Journal
J M Crane, A S Verkman

Abstract

The shorter "M23" isoform of the glial cell water channel aquaporin-4 (AQP4) assembles into orthogonal arrays of particles (OAPs) in cell plasma membranes, whereas the full-length "M1" isoform does not. N-terminal residues are responsible for OAP formation by AQP4-M23 and for blocking of OAP formation in AQP4-M1. In investigating differences in OAP formation by certain N-terminus mutants of AQP4, as measured by freeze-fracture electron microscopy versus live-cell imaging, we discovered reversible, temperature-dependent OAP assembly of certain weakly associating AQP4 mutants. Single-particle tracking of quantum-dot-labeled AQP4 in live cells and total internal reflection fluorescence microscopy showed >80% of M23 in OAPs at 10-50 degrees C compared to <10% of M1. However, OAP formation by N-terminus cysteine-substitution mutants of M1, which probe palmitoylation-regulated OAP assembly, was strongly temperature-dependent, increasing from <10% at 37 degrees C to >70% at 10 degrees C for the double mutant M1-C13A/C17A. OAP assembly by this mutant, but not by native M23, could also be modulated by reducing its membrane density. Exposure of native M1 and single cysteine mutants to 2-bromopalmitate confirmed the presence of regulated ...Continue Reading

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Citations

May 10, 2011·Acta Pharmacologica Sinica·Alan S VerkmanLukmanee Tradtrantip
Jul 29, 2010·The Journal of Biological Chemistry·Gunhild M MuellerRebecca P Hughey
Jan 11, 2011·The Journal of Biological Chemistry·Michael J Shipston
May 14, 2014·The Journal of General Physiology·Michael J Shipston
Sep 27, 2012·Trends in Parasitology·Matthew L JonesJulian C Rayner
Jun 21, 2012·The International Journal of Biochemistry & Cell Biology·Julien Ratelade, A S Verkman
Aug 17, 2010·Journal of Molecular Biology·Tadanori MitsumaYoshinori Fujiyoshi
Oct 15, 2013·Brain Pathology·A S VerkmanLukmanee Tradtrantip
Jun 19, 2013·The Biochemical Journal·Jungo KatoMasato Yasui
Jul 16, 2016·International Journal of Molecular Sciences·Giuseppe Felice MangiatordiOrazio Nicolotti
Feb 6, 2017·The Journal of Biological Chemistry·Anindit MukherjeeRebecca P Hughey

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