Molecular basis for carrier protein-dependent amide bond formation in the biosynthesis of lincosamide antibiotics

0573513 - MBÚ 2024 RIV DE eng J - Journal Article
Mori, T. - Kadlčík, Stanislav - Lyu, S. - Kameník, Zdeněk - Sakurada, K. - Mazumdar, Aninda - Wang, H. - Janata, Jiří - Abe, I.
Molecular basis for carrier protein-dependent amide bond formation in the biosynthesis of lincosamide antibiotics.
NAT CATAL. Roč. 6, č. 6 (2023), s. 531-542. ISSN 2520-1158. E-ISSN 2520-1158
R&D Projects: GA MŠMT(CZ) LX22NPO5103; GA ČR(CZ) GJ20-09811Y
Grant - others:AV ČR(CZ) LQ200202002
Program: Prémie Lumina quaeruntur
Institutional support: RVO:61388971
Keywords : phytochelatin-synthase * peptide * enzyme * lincomycin * synthetase * mechanism * papain * state * celesticetin * ligases
OECD category: Microbiology
Impact factor: 37.8, year: 2022
Method of publishing: Open access
https://www.nature.com/articles/s41929-023-00971-y

In the biosynthesis of the lincosamide antibiotic celesticetin, the condensation enzyme CcbD generates the lincosamide pharmacophore by forming an amide bond between the carrier protein (CP)-tethered proline and ergothioneine-conjugated thiooctose. Although the function of CcbD has been investigated, its substrate specificity, structures and catalytic mechanisms remain unclear. Here we show the structure–function analyses of CcbD. Our biochemical analysis revealed that CcbD exhibits promiscuous substrate specificity towards CP-tethered acyl substrates to generate unnatural lincosamides. Furthermore, structural analyses indicated that CcbD possesses an unusual overall fold, while the N-terminal region shows weak similarity to cysteine proteases. Thus, CcbD, like cysteine proteases, utilizes the Cys-His-Glu catalytic triad to form amide bonds in a CP-dependent manner, which is significantly different from other known amide bond-forming enzymes. Furthermore, the structures of the CcbD/thiooctose complex and the cross-linked CcbD/CcbZ-CP complex, as well as structure-based mutagenesis, revealed the intimate structural details of the CP-dependent amide bond formation reaction. [Figure not available: see fulltext.]
Permanent Link: https://hdl.handle.net/11104/0343947