Structure and Function of a C–C Bond Cleaving Oxygenase in Atypical Angucycline Biosynthesis

• DOI: 10.1021/acschembio.6b00621
• Publication Date: 2016-11-18T18:25:41Z
• Authors: Guohui Pan; Xiaoqin Gao; Keqiang Fan; Junlin Liu; Bing Meng; Jinmin Gao; Bin Wang; Chaobo Zhang; Hui Han; Guomin Ai; Yihua Chen; Dong Wu; Zhi-Jie Liu; Keqian Yang
• Keywords: Models, Molecular; 0303 health sciences; 03 medical and health sciences; Protein Domains; Protein Conformation; Polyketides; Oxygenases; Amino Acid Sequence; Sequence Alignment; Phylogeny; Streptomyces

C-C bond ring cleaving oxygenases represent a unique family of enzymes involved in the B cleavage reaction only observed atypical angucycline biosynthesis. is key leading to dramatic divergence final structures angucyclines. Here, we present crystal structure AlpJ, first this enzymes. AlpJ has been verified as enzyme catalyzing kinamycin The monomer resembles dimeric ferredoxin-like proteins. N- and C-terminal halves are homologous, both contain putative hydrophobic substrate binding pocket "closed" "open" conformations, respectively. Structural comparison with ActVA-Orf6 protein-ligand docking analysis suggest that residues including Asn60, Trp64, Trp181 possibly recognition. Site-directed mutagenesis results supported our hypothesis, mutation these led nearly complete loss activity AlpJ. also revealed possesses an intramolecular domain-domain interface, where His50 Tyr178 form hydrogen probably stabilizes three-dimensional showed two residues, Tyr178, were vital for Our findings shed light on catalytic mechanism oxygenases, which presumably involves active sites might function cooperative manner.