Experimental and computational investigation of enzyme functional annotations uncovers misannotation in the EC 1.1.3.15 enzyme class
Fig 3
Characterisation of protein clusters with low sequence identity to previously characterised S-2-hydroxyacid oxidases.
Dendrogram indicating protein relatedness. Superkingdoms: light purple—Bacteria, dark purple—Archaea. Activities are marked with squares; for proteins active with more than one substrate, the substrate preference is shaded. The cartoons represent predicted domain and motif composition of the sequences, based on Pfam search. Domains lacking full Pfam alignment are represented with a sharp edge. Proteins with alternative activities chosen for kinetic characterisation are marked in bold. (A) Characterisation of protein clusters containing DAO domain. FAD dependent oxidoreductase domain (DAO, PF01266) is marked in blue, BFD-like [2Fe-2S] binding domain (Fer2_BFD, PF04324) is marked in purple. (B) Characterisation of remaining protein clusters. FAD binding domain (FAD_binding_4, PF01565) is marked in orange, FAD linked oxidases C-terminal domain (FAD-oxidase_C, PF02913) is marked in green, cysteine rich domain (CCG, PF02754) is marked in red. (C) Comparison of Pfam domains of sequences annotated to EC 1.1.3.15 in BRENDA version 2017.1 and 2019.2.