Figure 1.

Nicotiana tabacum phytaspase (NtPhyt)-TurboID and signal peptide (SP)-TurboID proteins utilized for the identification of phytaspase interactors. (a) Schematic representation of recombinant proteins. Non-specific biotin ligase TurboID fused to SP derived from NtPhyt served as a control for in planta biotinylation accomplished by NtPhyt-TurboID protein. Both proteins bear His tags to permit their detection using Western blotting analysis. (b) Transient production of NtPhyt-TurboID and SP-TurboID in N. benthamiana leaves. Protein extracts from NtPhyt-TurboID-producing, SP-TurboID-producing and control (vector) leaves were fractionated by sodium dodecyl sulphate (SDS) gel-electrophoresis and analysed by Western blotting with HisProbe detection. Positions of molecular weight protein markers are indicated on the right.