8JZ5

Crystal structure of AetF in complex with FAD and NADP+ at 1.86 angstrom

Summary for 8JZ5

• Entry DOI: 10.2210/pdb8jz5/pdb
• Descriptor: AetF, FLAVIN-ADENINE DINUCLEOTIDE, NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, ... (4 entities in total)
• Functional Keywords: halogenase, flavin-dependent, single-component, nadp+, flavoprotein
• Biological source: Aetokthonos hydrillicola Thurmond2011
• Total number of polymer chains: 1
• Total formula weight: 80085.28

Authors

Li, H.,Dai, L.,Chen, C.-C.,Guo, R.-T. (deposition date: 2023-07-04, release date: 2024-01-17, Last modification date: 2024-06-05)

Primary citation

Dai, L.,Li, H.,Dai, S.,Zhang, Q.,Zheng, H.,Hu, Y.,Guo, R.T.,Chen, C.C.
Structural and functional insights into the self-sufficient flavin-dependent halogenase.
Int.J.Biol.Macromol., 260:129312-129312, 2024

PubMed Abstract: Flavin-dependent halogenases (FDHs) have tremendous applications in synthetic chemistry. A single-component FDH, AetF, exhibits both halogenase and reductase activities in a continuous polypeptide chain. AetF exhibits broad substrate promiscuity and catalyzes the two-step bromination of l-tryptophan (l-Trp) to produce 5-bromotryptophan (5-Br-Trp) and 5,7-dibromo-l-tryptophan (5,7-di-Br-Trp). To elucidate the mechanism of action of AetF, we solved its crystal structure in complex with FAD, FAD/NADP, FAD/l-Trp, and FAD/5-Br-Trp at resolutions of 1.92-2.23 Å. The obtained crystal structures depict the unprecedented topology of single-component FDH. Structural analysis revealed that the substrate flexibility and dibromination capability of AetF could be attributed to its spacious substrate-binding pocket. In addition, highly-regulated interaction networks between the substrate-recognizing residues and 5-Br-Trp are crucial for the dibromination activity of AetF. Several Ala variants underwent monobromination with >98 % C5-regioselectivity toward l-Trp. These results reveal the catalytic mechanism of single-component FDH for the first time and contribute to efficient FDH protein engineering for biocatalytic halogenation.

PubMed: 38216020
DOI: 10.1016/j.ijbiomac.2024.129312

Experimental method

X-RAY DIFFRACTION (1.86 Å)