8XFH

Crystal structure of MiCGT(E152Q/V190D/S122P) in complex with UDP

Summary for 8XFH

• Entry DOI: 10.2210/pdb8xfh/pdb
• Descriptor: UDP-glycosyltransferase 13, URIDINE-5'-DIPHOSPHATE (2 entities in total)
• Functional Keywords: transferase
• Biological source: Mangifera indica (mango)
• Total number of polymer chains: 2
• Total formula weight: 103640.17

Authors

Zhang, Z.M.,Zhou, Z.Q. (deposition date: 2023-12-13, release date: 2024-12-18, Last modification date: 2025-07-09)

Primary citation

Li, M.,Zhou, Y.,Wen, Z.,Ni, Q.,Zhou, Z.,Liu, Y.,Zhou, Q.,Jia, Z.,Guo, B.,Ma, Y.,Chen, B.,Zhang, Z.M.,Wang, J.B.
An efficient C-glycoside production platform enabled by rationally tuning the chemoselectivity of glycosyltransferases.
Nat Commun, 15:8893-8893, 2024

PubMed Abstract: Despite the broad potential applications of C-glycosides, facile synthetic methods remain scarce. Transforming glycosyltransferases with promiscuous or natural O-specific chemoselectivity to C-glycosyltransferases is challenging. Here, we employ rational directed evolution of the glycosyltransferase MiCGT to generate MiCGT-QDP and MiCGT-ATD mutants which either enhance C-glycosylation or switch to O-glycosylation, respectively. Structural analysis and computational simulations reveal that substrate binding mode govern C-/O-glycosylation selectivity. Notably, directed evolution and mechanism analysis pinpoint the crucial residues dictating the binding mode, enabling the rational design of four enzymes with superior non-inherent chemoselectivity, despite limited sequence homology. Moreover, our best mutants undergo testing with 34 substrates, demonstrating superb chemoselectivities, regioselectivities, and activities. Remarkably, three C-glycosides and an O-glycoside are produced on a gram scale, demonstrating practical utility. This work establishes a highly selective platform for diverse glycosides, and offers a practical strategy for creating various types of glycosylation platforms to access pharmaceutically and medicinally interesting products.

PubMed: 39406733
DOI: 10.1038/s41467-024-53209-1

Experimental method

X-RAY DIFFRACTION (2.80000300155 Å)