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ARS Home » Pacific West Area » Parlier, California » San Joaquin Valley Agricultural Sciences Center » Crop Diseases, Pests and Genetics Research » Research » Publications at this Location » Publication #411986
Research Project: Identifying Vulnerabilities in Vector-host-pathogen Interactions of Grapevine and Citrus Pathosystems to Advance Sustainable Management Strategies

Location: Crop Diseases, Pests and Genetics Research

Title: Protocol for quantitative evaluation of misfolded protein degradation using Agrobacterium-mediated expression system in Nicotiana benthamiana

Author
item CHENG, YANG - Nanjing Agricultural University
item MENG, RUI - Nanjing Agricultural University
item NIU, SIQI - Nanjing Agricultural University
item Peng, Hao
item 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). Article 103034. 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.