title: Fe(II)-dependent, uridine-5'-monophosphate α-ketoglutarate dioxygenases in the synthesis of 5'-modified nucleosides.
abstract: Several nucleoside antibiotics from various actinomycetes contain a high-carbon sugar nucleoside that is putatively derived via C-5'-modification of the canonical nucleoside. Two prominent examples are the 5'-C-carbamoyluridine- and 5'-C-glycyluridine-containing nucleosides, both families of which were discovered using screens aimed at finding inhibitors of bacterial translocase I involved in the assembly of the bacterial peptidoglycan cell wall. A shared open reading frame was identified whose gene product is similar to enzymes of the nonheme, Fe(II)-, and α-ketoglutarate-dependent dioxygenases. The enzyme LipL from the biosynthetic pathway for A-90289, a 5'-C-glycyluridine-containing nucleoside, was functionally characterized as an UMP:α-ketoglutarate dioxygenase, providing the enzymatic imperative for the generation of a nucleoside-5'-aldehdye that serves as a downstream substrate for an aldol or aldol-type reaction leading to the high-carbon sugar scaffold. The functional assignment of LipL and the homologous enzymes-including bioinformatic analysis, iron detection and quantification, and assay development for biochemical characterization-is presented herein.
authors.affiliation: Key Laboratory of Biotechnology of Antibiotics, Ministry of Health, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, PR China.
authors.forename: Zhaoyong
authors.initials: Z
authors.lastname: Yang
authors.forename: Jason
authors.initials: J
authors.lastname: Unrine
authors.forename: Koichi
authors.initials: K
authors.lastname: Nonaka
authors.forename: Steven G
authors.initials: SG
authors.lastname: Van Lanen
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002473:Cell Wall / Q000737:chemistry / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D019295:Computational Biology; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D007501:Iron / Q000737:chemistry / Q000378:metabolism; D007656:Ketoglutaric Acids / Q000737:chemistry / Q000378:metabolism*; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010457:Peptidoglycan / Q000737:chemistry / Q000378:metabolism; D013302:Streptomyces / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D014542:Uridine Monophosphate / Q000031:analogs & derivatives / Q000096:biosynthesis* / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D007656:Ketoglutaric Acids; D010457:Peptidoglycan; D007501:Iron; D014542:Uridine Monophosphate; D006899:Mixed Function Oxygenases; D008049:Lipase; C120013:lactonizing lipase protein, Pseudomonas