8HOJ

Crystal structure of UGT71AP2 in complex with UDP

Summary for 8HOJ

• Entry DOI: 10.2210/pdb8hoj/pdb
• Descriptor: UGT71AP2, URIDINE-5'-DIPHOSPHATE (2 entities in total)
• Functional Keywords: glycosyltransferase, transferase
• Biological source: Scutellaria baicalensis
• Total number of polymer chains: 2
• Total formula weight: 104040.32

Authors

Wang, Z.L.,He, C.,Li, F.,Qiao, X.,Ye, M. (deposition date: 2022-12-10, release date: 2023-12-13, Last modification date: 2024-09-18)

Primary citation

Wang, Z.,Du, X.,Ye, G.,Wang, H.,Liu, Y.,Liu, C.,Li, F.,Agren, H.,Zhou, Y.,Li, J.,He, C.,Guo, D.A.,Ye, M.
Functional characterization, structural basis, and protein engineering of a rare flavonoid 2'- O -glycosyltransferase from Scutellaria baicalensis .
Acta Pharm Sin B, 14:3746-3759, 2024

PubMed Abstract: Glycosylation is an important post-modification reaction in plant secondary metabolism, and contributes to structural diversity of bioactive natural products. In plants, glycosylation is usually catalyzed by UDP-glycosyltransferases. Flavonoid 2'--glycosides are rare glycosides. However, no UGTs have been reported, thus far, to specifically catalyze 2'--glycosylation of flavonoids. In this work, UGT71AP2 was identified from the medicinal plant as the first flavonoid 2'--glycosyltransferase. It could preferentially transfer a glycosyl moiety to 2'-hydroxy of at least nine flavonoids to yield six new compounds. Some of the 2'--glycosides showed noticeable inhibitory activities against cyclooxygenase 2. The crystal structure of UGT71AP2 (2.15 Å) was solved, and mechanisms of its regio-selectivity was interpreted by p calculations, molecular docking, MD simulation, MM/GBSA binding free energy, QM/MM, and hydrogen‒deuterium exchange mass spectrometry analysis. Through structure-guided rational design, we obtained the L138T/V179D/M180T mutant with remarkably enhanced regio-selectivity (the ratio of 7--glycosylation byproducts decreased from 48% to 4%) and catalytic efficiency of 2'--glycosylation ( / , 0.23 L/(s·μmol), 12-fold higher than the native). Moreover, UGT71AP2 also possesses moderate UDP-dependent de-glycosylation activity, and is a dual function glycosyltransferase. This work provides an efficient biocatalyst and sets a good example for protein engineering to optimize enzyme catalytic features through rational design.

PubMed: 39220864
DOI: 10.1016/j.apsb.2024.04.001

Experimental method

X-RAY DIFFRACTION (2.9 Å)