Location: Crop Diseases, Pests and Genetics Research
Title: Protocol for quantitative evaluation of misfolded protein degradation using Agrobacterium-mediated expression system in Nicotiana benthamianaAuthor
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CHENG, YANG - Nanjing Agricultural University |
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MENG, RUI - Nanjing Agricultural University |
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NIU, SIQI - Nanjing Agricultural University |
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Peng, Hao |
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JING, MAOFENG - Nanjing Agricultural University |
Submitted to: STAR (Structured Transparent Accessible Reproducible) Protocols
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 4/10/2024 Publication Date: 4/24/2024 Citation: Cheng, Y., Meng, R., Niu, S., Peng, H., Jing, M. 2024. Protocol for quantitative evaluation of misfolded protein degradation using Agrobacterium-mediated expression system in Nicotiana benthamiana. STAR (Structured Transparent Accessible Reproducible) Protocols. 5(2). https://doi.org/10.1016/j.xpro.2024.103034. DOI: https://doi.org/10.1016/j.xpro.2024.103034 Interpretive Summary: When proteins are folded wrongly in the cell, they will be degraded. Here, we develop a quantitative protocol to evaluate the degradation of misfolded proteins using Agrobacterium-mediated expression system in Nicotiana benthamiana (a close relative of tobacco). This protocol is useful for examining the degradation mechanisms of misfolded proteins from various plants or microbes. Technical Abstract: Cells must dispose of aggregated misfolded proteins in order to maintain cellular homeostasis. Here, we present a protocol to investigate the role of target protein in degrading misfolded protein via Nicotiana benthamiana expression system. We describe the steps for design of misfolded proteins, Agrobacterium-mediated transient expression in N. benthamiana. We then detail the extraction of total proteins and their quantification of misfolded proteins through western blotting. This system is generalizable and can be used for various plants or microbes. |