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. 2019 Sep 27;10:1164. doi: 10.3389/fpls.2019.01164

Figure 5.

Figure 5

Glycosyltransferase assay of UGT76C4, UGT76C5, and their mutants. (A) Protein sequence alignment of UGT76C4 and UGT76C5. The PSPG motif was boxed in green, and the different amino acids in PSPG motif between UGT76C4 and UGT76C5 were indicated with black arrows and numbers ① (D340 in UGT76C5 or E336 in UGT76C4), ② (S357 in UGT76C5 or N353 in UGT76C4), and ③ (A366 in UGT76C5 or G362 in UGT76C4). Among 44 amino acids in PSPG motif, the amino acids marked with red number are putatively bound to UDP-sugars. (B) Biochemical assay of UGT76C4, UGT76C5, and their mutants with different sugar donors and [14C]-NA. N.C., negative control; only purified MBP tag protein was added in this reaction. W.T., wild-type. ①, amino acid interconversion between D and E; ②, amino acid interconversion between S and N; and ③, amino acid interconversion between A and G. (C) Comparisons of glycosyltransferase activity of UGT76C4 and UGT76C5 wild-type and mutant enzymes with various UDP-sugars. W.T., wild type. Bars show means ± SDs (n = 3).