Fig. 4: Mutational analysis of interactions between CtAms1 and CtNbr1-FW.
From: Structural mechanism of protein recognition by the FW domain of autophagy receptor Nbr1
a–c Pil1 co-tethering assay. CtNbr1 FW domain and its mutants were fused to Pil1-mCherry and co-expressed with CtAms1-mECitrine or its mutants in S. pombe. Cells were imaged by fluorescence microscopy. Bar, 3 μm. The data shown are representative of two independent experiments with similar results. Source data are provided as a Source Data file. d SPR analysis of the CtNbr1-CtAms1 interaction. GST-tagged CtNbr1 FW domain or its mutants was bound to a CM5 sensor chip. CtAms1 or CtAms1-G2A (twofold serial dilutions from 50 nM to 1.56 nM) flowed through the chip surface. The experiments were performed once. e SPR analysis of the interactions of CtNbr1 and hNBR1 with CtAms1 N-terminal peptides. GST-tagged CtNbr1 FW domain or hNBR1 FW domain was bound to a CM5 sensor chip. Chemically synthesized CtAms1 N-terminal peptides (twofold serial dilutions from 100 μM to 3.13 μM) flowed through the chip surface. The experiments were performed twice with similar results.
