Structure-Function Analysis of the S-Glycosylation Reaction in the Biosynthesis of Lincosamide Antibiotics

0573702 - MBÚ 2024 RIV DE eng J - Journal Article
Mori, T. - Sun, X. - Kadlčík, Stanislav - Janata, Jiří - Abe, I.
Structure-Function Analysis of the S-Glycosylation Reaction in the Biosynthesis of Lincosamide Antibiotics.
Angewandte Chemie - International Edition. Roč. 62, č. 29 (2023), č. článku e2023049. ISSN 1433-7851. E-ISSN 1521-3773
R&D Projects: GA ČR(CZ) GJ20-09811Y; GA MŠMT(CZ) LX22NPO5103
Grant - others:AV ČR(CZ) LQ200202002
Program: Prémie Lumina quaeruntur
Institutional support: RVO:61388971
Keywords : LmbT * S-glycosyltransferase * lincomycin * ergothioneine * protein structure
OECD category: Microbiology
Impact factor: 16.6, year: 2022
Method of publishing: Open access
https://onlinelibrary.wiley.com/doi/10.1002/anie.202304989

The S-glycosyltransferase LmbT, involved in the biosynthesis of lincomycin A, is the only known enzyme that catalyzes the enzymatic incorporation of rare amino acid L-ergothioneine (EGT) into secondary metabolites. Here, we show the structure and function analyses of LmbT. Our in vitro analysis of LmbT revealed that the enzyme shows promiscuous substrate specificity toward nitrogenous base moieties in the generation of unnatural nucleotide diphosphate (NDP)-D-α-D-lincosamides. Furthermore, the X-ray crystal structures of LmbT in its apo form and in complex with substrates indicated that the large conformational changes of the active site occur upon binding of the substrates, and that EGT is strictly recognized by salt-bridge and cation-π interactions with Arg260 and Trp101, respectively. The structure of LmbT in complex with its substrates, the docking model with the EGT-S-conjugated lincosamide, and the structure-based site-directed mutagenesis analysis revealed the structural details of the LmbT-catalyzed SN2-like S-glycosylation reaction with EGT.
Permanent Link: https://hdl.handle.net/11104/0344076