Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/137713
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Type: Journal article
Title: A high-throughput yeast approach to characterize aquaporin permeabilities: Profiling the Arabidopsis PIP aquaporin sub-family
Author: Groszmann, M.
De Rosa, A.
Chen, W.
Qiu, J.
McGaughey, S.A.
Byrt, C.S.
Evans, J.R.
Citation: Frontiers in Plant Science, 2023; 14:1078220-1-1078220-19
Publisher: Frontiers Media
Issue Date: 2023
ISSN: 1664-462X
1664-462X
Statement of
Responsibility: 
Michael Groszmann, Annamaria De Rosa, Weihua Chen, Jiaen Qiu, Samantha A. McGaughey, Caitlin S. Byrt and John R. Evans
Abstract: Introduction: Engineering membrane transporters to achieve desired functionality is reliant on availability of experimental data informing structure-function relationships and intelligent design. Plant aquaporin (AQP) isoforms are capable of transporting diverse substrates such as signaling molecules, nutrients, metalloids, and gases, as well as water. AQPs can act as multifunctional channels and their transport function is reliant on many factors, with few studies having assessed transport function of specific isoforms for multiple substrates. Methods: High-throughput yeast assays were developed to screen for transport function of plant AQPs, providing a platform for fast data generation and cataloguing of substrate transport profiles.We applied our high-throughput growth-based yeast assays to screen all 13 Arabidopsis PIPs (AtPIPs) for transport of water and several neutral solutes: hydrogen peroxide (H2O2), boric acid (BA), and urea. Sodium (Na+) transport was assessed using elemental analysis techniques. Results: All AtPIPs facilitated water and H2O2 transport, although their growth phenotypes varied, and none were candidates for urea transport. For BA and Na+ transport, AtPIP2;2 and AtPIP2;7 were the top candidates, with yeast expressing these isoforms having the most pronounced toxicity response to BA exposure and accumulating the highest amounts of Na+. Linking putative AtPIP isoform substrate transport profiles with phylogenetics and gene expression data, enabled us to align possible substrate preferences with known and hypothesized biological roles of AtPIPs. Discussion: This testing framework enables efficient cataloguing of putative transport functionality of diverse AQPs at a scale that can help accelerate our understanding of AQPbiology through big data approaches (e.g.association studies).The principles of the individual assays could be further adapted to test additional substrates. Data generated from this framework could inform future testing of AQP physiological roles, and address knowledge gaps in structure-function relationships to improve engineering efforts.
Keywords: aquaporin (AQP)
PIP
membrane channel proteins
high-throughput (HT) screening
heterologous yeast expression
protein engineering
Description: PUBLISHED 19 January 2023
Rights: © 2023 Groszmann, De Rosa, Chen, Qiu, McGaughey, Byrt and Evans. This is an openaccess article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
DOI: 10.3389/fpls.2023.1078220
Grant ID: http://purl.org/au-research/grants/arc/CE140100015
http://purl.org/au-research/grants/arc/DP190102725
http://purl.org/au-research/grants/arc/FT180100476
http://purl.org/au-research/grants/arc/CE140100008
Published version: http://dx.doi.org/10.3389/fpls.2023.1078220
Appears in Collections:Agriculture, Food and Wine publications

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