Fig. 3
Characteristics of PbrADC1. a PbrADC1 self-interaction determination. (a-i) Y2H assay. Transformants containing AD-T & BD-53, AD-T & BD-Lam, AD & BD, AD & BD-PbrADC1, and AD-PbrADC1 & BD were used as controls. (a-ii) BiFC assay. Transformants containing YFPN & YFPC, YFPN & PbrADC1-YFPC, and YFPC & PbrADC1-YFPN were used as controls. b Identification and functional validation of the substrate-binding residue Cys546 in PbrADC1. (b-i) Alignment of PbrADC1 with AtADCs and HpADC1 by Jalview Version 2. Information on AtADCs and HpADC1 was summarized by Hanfrey et al. (2001). (b-ii) Functional validation of Cys546. Enzyme activities of the His-tagged recombinant PbrADC1 and PbrADC1.C546A proteins were analyzed based on the method of Song et al. (2010). c Exogenous H2O2 treatment on PbrADC1 activity. (c-i) Identification of the H2O2-modified Cys residues in PbrADC1. The H2O2-modified Cys residues in PbrADC1 was predicted by pCysMod database (Li et al. 2021b). (c-ii) Impact of exogenous H2O2 treatment on PbrADC1 activity. The residual activity was expressed as a percentage of the initial (0 min). Data represented the mean value of three biological replicates, and different lowercase letters meant significance between samples (p < 0.05)