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Screening of Proximal and Interacting Proteins in Rice Protoplasts by Proximity-Dependent Biotinylation

Proximity-dependent biotin identification (BioID), which detects physiologically relevant proteins based on the proximity-dependent biotinylation process, has been successfully used in different organisms. In this report, we established the BioID system in rice protoplasts. Biotin ligase BirAG was o...

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Autores principales: Lin, Qiupeng, Zhou, Zejiao, Luo, Wanbin, Fang, Maichun, Li, Meiru, Li, Hongqing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5427108/
https://www.ncbi.nlm.nih.gov/pubmed/28553299
http://dx.doi.org/10.3389/fpls.2017.00749
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author Lin, Qiupeng
Zhou, Zejiao
Luo, Wanbin
Fang, Maichun
Li, Meiru
Li, Hongqing
author_facet Lin, Qiupeng
Zhou, Zejiao
Luo, Wanbin
Fang, Maichun
Li, Meiru
Li, Hongqing
author_sort Lin, Qiupeng
collection PubMed
description Proximity-dependent biotin identification (BioID), which detects physiologically relevant proteins based on the proximity-dependent biotinylation process, has been successfully used in different organisms. In this report, we established the BioID system in rice protoplasts. Biotin ligase BirAG was obtained by removing a cryptic intron site in the BirA(∗) gene when expressed in rice protoplasts. We found that protein biotinylation in rice protoplasts increased with increased expression levels of BirAG. The biotinylation effects can also be achieved by exogenous supplementation of high concentrations of biotin and long incubation time with protoplasts. By using this system, multiple proteins were identified that associated with and/or were proximate to OsFD2 in vivo. Our results suggest that BioID is a useful and generally applicable method to screen for both interacting and neighboring proteins in their native cellular environment in plant cell.
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spelling pubmed-54271082017-05-26 Screening of Proximal and Interacting Proteins in Rice Protoplasts by Proximity-Dependent Biotinylation Lin, Qiupeng Zhou, Zejiao Luo, Wanbin Fang, Maichun Li, Meiru Li, Hongqing Front Plant Sci Plant Science Proximity-dependent biotin identification (BioID), which detects physiologically relevant proteins based on the proximity-dependent biotinylation process, has been successfully used in different organisms. In this report, we established the BioID system in rice protoplasts. Biotin ligase BirAG was obtained by removing a cryptic intron site in the BirA(∗) gene when expressed in rice protoplasts. We found that protein biotinylation in rice protoplasts increased with increased expression levels of BirAG. The biotinylation effects can also be achieved by exogenous supplementation of high concentrations of biotin and long incubation time with protoplasts. By using this system, multiple proteins were identified that associated with and/or were proximate to OsFD2 in vivo. Our results suggest that BioID is a useful and generally applicable method to screen for both interacting and neighboring proteins in their native cellular environment in plant cell. Frontiers Media S.A. 2017-05-12 /pmc/articles/PMC5427108/ /pubmed/28553299 http://dx.doi.org/10.3389/fpls.2017.00749 Text en Copyright © 2017 Lin, Zhou, Luo, Fang, Li and Li. http://creativecommons.org/licenses/by/4.0/ This is an open-access 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) or licensor 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.
spellingShingle Plant Science
Lin, Qiupeng
Zhou, Zejiao
Luo, Wanbin
Fang, Maichun
Li, Meiru
Li, Hongqing
Screening of Proximal and Interacting Proteins in Rice Protoplasts by Proximity-Dependent Biotinylation
title Screening of Proximal and Interacting Proteins in Rice Protoplasts by Proximity-Dependent Biotinylation
title_full Screening of Proximal and Interacting Proteins in Rice Protoplasts by Proximity-Dependent Biotinylation
title_fullStr Screening of Proximal and Interacting Proteins in Rice Protoplasts by Proximity-Dependent Biotinylation
title_full_unstemmed Screening of Proximal and Interacting Proteins in Rice Protoplasts by Proximity-Dependent Biotinylation
title_short Screening of Proximal and Interacting Proteins in Rice Protoplasts by Proximity-Dependent Biotinylation
title_sort screening of proximal and interacting proteins in rice protoplasts by proximity-dependent biotinylation
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5427108/
https://www.ncbi.nlm.nih.gov/pubmed/28553299
http://dx.doi.org/10.3389/fpls.2017.00749
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