Fig. 2: Exploring the mechanism of unique tyrosine specificity of NnSULT1C1. | Nature Communications

Fig. 2: Exploring the mechanism of unique tyrosine specificity of NnSULT1C1.

From: Unleashing the potential of noncanonical amino acid biosynthesis to create cells with precision tyrosine sulfation

Fig. 2

a NnSULT1C1 structure (blue) predicted by AlphaFold2 and its active site consisting PAPS and Tyr. Tyr was docked into NnSULT1C1 containing PAPS by Glide v8.1 in Schrödinger software. b Green fluorescent protein assay with wildtype NnSULT1C1 (wt) or NnSULT1C1 without the SIQEPPAAS (Δloop), p = 0.01. c Green fluorescent protein assay with wildtype NnSULT1C1 (wt) or NnSULT1C1 with alanine mutation at indicated resiudes, p = 0.004. d Structural similarity search of NnSULT1C1 using the PDBeFold web server. e Characterization of Tyr docking with NnSULT1C1 and its structurally similar sulfotransferases via docking score and nucleophilic attack distance. Docking scores were calculated using Glide v8.1 in Schrödinger software. Nucleophilic attack distance was defined as the distances between Tyr phenolic alcohol and PAPS sulfonate. f Comparison of tyrosine sulfation activity of NnSULT1C1 and its structurally similar sulfotransferases using green fluorescent protein assay. Cells without any sulfotransferase (−) were used as control. g–j Tyr docking position with NnSULT1C1 (g), mSULT1D1 (h), hSULT1A3 (i), and hSULT1C2 (j). PAPS and Tyr are shown as sticks with green carbon. Docking was performed by Glide v8.1 in Schrödinger software with the same parameters in a. b, c, f Data are plotted as the mean ± standard deviation from n = 3 independent samples. b, c Two-sided unpaired t-tests were performed with *p < 0.05; **p < 0.01. a.u. stands for arbitrary unit.

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