TY - JOUR
T1 - Protocol to identify protein-protein interaction networks in Solanum tuberosum using transient TurboID-based proximity labeling
AU - Shi, Li
AU - Marti Ferrando, Tatiana
AU - Landeo Villanueva, Sergio
AU - Joosten, Matthieu H.A.J.
AU - Vleeshouwers, Vivianne G.A.A.
AU - Bachem, Christian W.B.
PY - 2023/12/15
Y1 - 2023/12/15
N2 - 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.
AB - 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.
KW - Molecular Biology
KW - Plant Sciences
KW - Protein Biochemistry
U2 - 10.1016/j.xpro.2023.102577
DO - 10.1016/j.xpro.2023.102577
M3 - Article
AN - SCOPUS:85171462863
SN - 2666-1667
VL - 4
JO - STAR Protocols
JF - STAR Protocols
IS - 4
M1 - 102577
ER -