Fig. 2

Sequence alignment and protein modelling analyses. a Amino acid sequence alignment of TaALP-ax-4AL proteins in CS, Spitefire, and Mace. Cysteine residues were highlighted in blue. Secondary structural elements based on protein modelling were displayed above the sequence alignment. Three predicted disulphate bonds were underlined in pink number 1, 2, and 3. b Superimposition of the tertiary structure models of TaALP-ax-4AL in CS (green) and Mace (cyan). Disulphate bonds were displayed in sticks (yellow). The amino acid substitution sites were displayed in red. Only a single substitution (S169 N) exists between CS and Spitfire. c Displays the hydrophobicity profile. The substitution site residues with hydrophobicity change were shown in sticks, with red and white colours indicating the most hydrophobic and the most hydrophilic residues, respectively. Protein models were generated using the I-TASSER server [64, 65]. Structure visualization was implemented in PyMol (v1.7.4.5)