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Updated: Jan 17, 2026

TurboID-Based Proximity Labeling for In Planta Identification of Protein-Protein Interaction Networks
Published on: May 17, 2020
Pupylation-based proximity labeling reveals APC/C-mediated ubiquitination and degradation of PEROXIN11C in soybean
Yue Zhang1, Xinxin Yang1, Tong Chen1
1The Key Laboratory of Plant Development and Environmental Adaptation Biology, Ministry of Education, Shandong Key Laboratory of Precision Molecular Crop Design and Breeding, School of Life Sciences, Shandong University, Qingdao 266237, China.
Abstract:
Proximity-dependent labeling coupled with mass spectrometry (PL-MS) is a powerful approach for mapping low-affinity, transient protein-protein interactions (PPIs). While widely employed in animal systems, its application in plants has been restricted primarily to TurboID/miniTurbo-based systems due to the limited enzyme compatibility. To address this, we optimized the pupylation-based interaction tagging (Pup-IT) with a dexamethasone-inducible expression strategy in Nicotiana benthamiana and soybean (Glycine max). Using Pup-IT, we mapped the interactome of the soybean anaphase-promoting complex/cyclosome (APC/C) and identified the peroxisomal membrane protein GmPEX11C as a substrate. GmAPC/C ubiquitinates GmPEX11C at lysine 116, targeting it for proteasomal degradation. In addition, Gmilpa1/apc8 mutant exhibited enhanced salt and drought tolerance, whereas knockdown of GmPEX11C via RNA interference abolishes this stress-resistant phenotype. Our study establishes Pup-IT as a useful tool for interrogating PPIs in planta and uncovers a mechanistic link between APC/C-mediated ubiquitination and abiotic stress adaptation in soybean, offering potential avenues for crop improvement.
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