Crystal Structure of a Yeast Aquaporin at 1.15 Ã//Reveals a Novel Gating Mechanism
Citation: Gerhard Fischer, Urszula Kosinska-Eriksson, Camilo Aponte-SantamarÃa, Madelene Palmgren, Cecilia Geijer, Kristina Hedfalk, Stefan Hohmann, Bert L. de Groot, Richard Neutze, Karin Lindkvist-Petersson (2009/06) Crystal Structure of a Yeast Aquaporin at 1.15 Ã//Reveals a Novel Gating Mechanism. PLoS Biol (Volume 7) (RSS)
DOI (original publisher): 10.1371/journal.pbio.1000130
Semantic Scholar (metadata): 10.1371/journal.pbio.1000130
Sci-Hub (fulltext): 10.1371/journal.pbio.1000130
Internet Archive Scholar (search for fulltext): Crystal Structure of a Yeast Aquaporin at 1.15 Ã//Reveals a Novel Gating Mechanism
Download: http://dx.doi.org/10.1371/journal.pbio.1000130
Tagged: Biology
(RSS) aquaporins (RSS)
Summary
Aquaporins are transmembrane proteins that facilitate the flow of water through cellular membranes. An unusual characteristic of yeast aquaporins is that they frequently contain an extended N terminus of unknown function. Here we present the X-ray structure of the yeast aquaporin Aqy1 from Pichia pastoris at 1.15 resolution. Our crystal structure reveals that the water channel is closed by the N terminus, which arranges as a tightly wound helical bundle, with Tyr31 forming H-bond interactions to a water molecule within the pore and thereby occluding the channel entrance. Nevertheless, functional assays show that Aqy1 has appreciable water transport activity that aids survival during rapid freezing of P. pastoris. These findings establish that Aqy1 is a gated water channel. Mutational studies in combination with molecular dynamics simulations imply that gating may be regulated by a combination of phosphorylation and mechanosensitivity.
All living organisms must regulate precisely the flow of water into and out of cells in order to maintain cell shape and integrity. Proteins of one family, the aquaporins, are found in virtually every living organism and play a major role in maintaining water homeostasis by acting as regulated water channels. Here we describe the first crystal structure of a yeast aquaporin, Aqy1, at 1.15 resolution, which represents the highest resolution structural data obtained to date for a membrane protein. Using this structural information, we address an outstanding biological question surrounding yeast aquaporins: what is the functional role of the amino-terminal extension that is characteristic of yeast aquaporins? Our structural data show that the amino terminus of Aqy1 fulfills a novel gate-like function by folding to form a cytoplasmic helical bundle with a tyrosine residue entering the water channel and occluding the cytoplasmic entrance. Molecular dynamics simulations and functional studies in combination with site-directed mutagenesis suggest that water flow is regulated through a combination of mechanosensitive gating and post-translational modifications such as phosphorylation. Our study therefore provides insight into a unique mechanism for the regulation of water flux in yeast.
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