Amino acid sequence alignment of Lotus japonicus CHIT5, Medicago truncatula NFH1 (KC833515) and Arabidopsis thaliana CHIC (NP_193716). (*) indicates a conserved residue, (:) indicates strongly similar properties, (.) indicates weakly similar properties, (-) indicates gaps. A predicted signal peptide is indicated in bold for CHIT5. The conserved catalytic motif DxDxE is indicated as is the position of the E-K mutation (green) introduced to produce an inactive CHIT5 construct. The position of the P-L substitution in chit5-2 is indicated (magenta). The position of A and B loops (in red) and of the proline residue (red box) identified as important for Nod factor hydrolase activity of MtNFH1 are indicated.