Additional file 1 of Revealing two important tryptophan residues with completely different roles in a dye-decolorizing peroxidase from Irpex lacteus F17
Creators: name: Li, Liuqing
name: Wang, Tao
name: Chen, Taohua
name: Huang, Wenhan
name: Zhang, Yinliang
name: Jia, Rong
name: He, Chao
Publisher: name: figshare
Year: 2021
Subjects: subject: Inorganic Chemistry
subject: FOS: Chemical sciences

description: Additional file 1: Table S1 The primers sequences used in this study. Table S2 The structures of substrates used in this study. Fig. S1 Sequence alignments of tryptophan and tyrosine residues in Il-DyP4 with other representative class V DyPs. Highlighted residues include the following: (i) tryptophan residues were shown in red and the corresponding tryptophans in Il-DyP4 were indicated in the black box. (ii) tyrosine residues were shown in green and the corresponding tyrosines in Il-DyP4 were indicated in the black box. Above amino acid sequences of class V DyPs were downloaded from NCBI database: DyP2 from Amycolatopsis sp. (GI: 496374264), AnaPX from Anabaena sp. (GI: 1772287408), FmDyP from Fomitiporia mediterranea (GI: 595785908), VvDyP from Volvariella volvacea (GenBank: AKU04643.1), TvDyP1 from Trametes versicolor (GI: 636616485), ShDyP from Stereum hirsutum (GI: 597911761), HiDyP from Heterobasidion irregulare (GI: 575067022), GlDyP from Ganoderma lucidum (GenBank: ADN05763.1), DsDyP from Dichomitus squalens (GI: 1585543766), AauDyP from Auricularia auricula-judae (GI: 1048348430) and BadDyP from Bjerkandera adusta (GI: 116666995). Fig. S2 Electronic absorbance spectra of Il-DyP4 and the variants. a Absorbance spectra of Il-DyP4 and the W variants. b Absorbance spectra of Il-DyP4 and the W380 variants. All samples used in these assays had about 0.1–0.11 mg/mL of each of enzymes in 10 mM sodium acetate buffer, pH 6.0. Fig. S3 The structure comparison between Il-DyP4 and other fungal DyPs. a-c represent Il-DyP4 (PDB: 7D8M), BadDyP from Bjerkandera adusta (PDB: 3AFV) and AauDyP from Auricularia auricula-judae (PDB: 4W7J), respectively. d–f show the important residues surrounding the heme of each protein and the suggested surface-exposed catalytic tryptophan is showed in orange. Fig. S4 The location of W264 in Il-DyP4 and the comparison with other fungal DyPs. a represents the distances (Å) between the above tryptophan residues and the heme iron in Il-DyP4; b-d represent Il-DyP4, BadDyP and AauDyP, respectively, showing the position of W264 in Il-DyP4 and the corresponding residues in BadDyP and AauDyP. Fig. S5 The structure comparison between W and those amino acids used in W264 mutation. Fig. S6 a shows the distance between W264 and H312. b shows the F261 between W264 and H312. Yellow sticks represent the heme cofactor. Fig. S7 Location of W380 in Il‑DyP4. The molecular surface is shown in white. The red sphere represents W380. Fig. S8 The hydrophobic phenylalanines surrounding W380 in Il‑DyP4. The yellow sticks represent the heme cofactor. Fig. S9 Structural superpositions of Il‑DyP4 (yellow, PDB: 7D8M) and AauDyP (azure, PDB: 4W7J) showing a similar electron transfer pathway from the surface-exposed W380 to heme (according to Linde et al. 2015 [27]).