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Protocol to identify protein-protein interaction networks in Solanum tuberosum using transient TurboID-based proximity labeling

Protein-protein interactions (PPIs) in crop plants remain largely unexplored. Here, we provide a protocol for identifying PPIs in potato (Solanum tuberosum) using TurboID-mediated proximity labeling. We transiently expressed constructs for a nucleus-located transcription factor and a plasma membrane...

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Detalles Bibliográficos
Autores principales: Shi, Li, Marti Ferrando, Tatiana, Landeo Villanueva, Sergio, Joosten, Matthieu H.A.J., Vleeshouwers, Vivianne G.A.A., Bachem, Christian W.B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10519850/
https://www.ncbi.nlm.nih.gov/pubmed/37733594
http://dx.doi.org/10.1016/j.xpro.2023.102577
Descripción
Sumario:Protein-protein interactions (PPIs) in crop plants remain largely unexplored. Here, we provide a protocol for identifying PPIs in potato (Solanum tuberosum) using TurboID-mediated proximity labeling. We transiently expressed constructs for a nucleus-located transcription factor and a plasma membrane-localized receptor-like kinase fused to TurboID to identify PPIs in potato leaves. We describe the plasmid construction, plant material, agroinfiltration, biotin treatment, protein isolation, free biotin removal, western blot analysis, and enrichment of biotinylated proteins for mass spectrometry analysis.