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Structural biology of auxin efflux carrier PIN proteins
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Zhou, Xinghao (2021) Structural biology of auxin efflux carrier PIN proteins. PhD thesis, University of Warwick.
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Official URL: http://webcat.warwick.ac.uk/record=b3857506
Abstract
Auxin is an important hormone in plants which regulates plant growth and development. The plant-specific proteins named PIN-FORMED auxin efflux carriers (PINs) control the direction of auxin flow and thus play a necessary role in the local auxin distribution within plant tissues and organs, and consequently guide plant ontogenesis. PINs are membrane proteins with two hydrophobic regions consisting of five transmembrane helices linked with a hydrophilic loop. Normally plasma membrane-localized PINs have longer loops then endoplasmic reticulum PINs. The PIN1 secondary structure was published in 1999, but the three-dimensional structure of PIN has not been solved yet.
This project aimed to express and purify PIN proteins for structural studies. 8 different PINs from 5 plant species were expressed in insect Sf9 cells and in Saccharomyces cerevisiae. A series of purification optimizations for PIN proteins were applied and Arabidopsis thaliana PIN5 and Oryza sativa PIN8 were purified. However, high purity combined with the high yields necessary for crystallography trails were not achieved, due to the instability of PINs. Some negative stain electron microscopy suggested a unit structure of a dimer, but at very low resolution and Saposin A nanodisc lipid complexes were investigated to try and improve PIN electron microscopy studies.
A range of approaches in this project have allowed us to learn more about PIN structure and hopefully a more comprehensive understanding of the PIN mechanism of action will be obtained in the future
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QK Botany Q Science > QP Physiology |
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Library of Congress Subject Headings (LCSH): | Auxin, Plants -- Effect of auxin on, Auxin -- Molecular aspects, Membrane proteins | ||||
Official Date: | November 2021 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | School of Life Sciences | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Napier, R. (Richard) ; Cameron, Alexander | ||||
Format of File: | |||||
Extent: | xii, 145 pages : illustrations | ||||
Language: | eng |
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