title: X-ray analysis of butirosin biosynthetic enzyme BtrN redefines structural motifs for AdoMet radical chemistry.
abstract: The 2-deoxy-scyllo-inosamine (DOIA) dehydrogenases are key enzymes in the biosynthesis of 2-deoxystreptamine-containing aminoglycoside antibiotics. In contrast to most DOIA dehydrogenases, which are NAD-dependent, the DOIA dehydrogenase from Bacillus circulans (BtrN) is an S-adenosyl-l-methionine (AdoMet) radical enzyme. To examine how BtrN employs AdoMet radical chemistry, we have determined its structure with AdoMet and substrate to 1.56 Å resolution. We find a previously undescribed modification to the core AdoMet radical fold: instead of the canonical (β/α)6 architecture, BtrN displays a (β5/α4) motif. We further find that an auxiliary [4Fe-4S] cluster in BtrN, thought to bind substrate, is instead implicated in substrate-radical oxidation. High structural homology in the auxiliary cluster binding region between BtrN, fellow AdoMet radical dehydrogenase anSME, and molybdenum cofactor biosynthetic enzyme MoaA provides support for the establishment of an AdoMet radical structural motif that is likely common to ~6,400 uncharacterized AdoMet radical enzymes.
authors.affiliation: Departments of Chemistry and Biology and Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Goldman
authors.forename: Tyler L
authors.initials: TL
authors.lastname: Grove
authors.forename: Squire J
authors.initials: SJ
authors.lastname: Booker
authors.forename: Catherine L
authors.initials: CL
authors.lastname: Drennan
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D053898:Biosynthetic Pathways / Q000235:genetics*; D002076:Butirosin Sulfate / Q000096:biosynthesis*; D002237:Carbohydrate Dehydrogenases / Q000737:chemistry* / Q000378:metabolism; D003460:Crystallization; D017931:DNA Primers / Q000235:genetics; D058085:Iron Compounds / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D011485:Protein Binding; D011487:Protein Conformation; D012436:S-Adenosylmethionine / Q000378:metabolism; D013457:Sulfur Compounds / Q000378:metabolism
chemical_list: D017931:DNA Primers; D058085:Iron Compounds; D013457:Sulfur Compounds; D002076:Butirosin Sulfate; D012436:S-Adenosylmethionine; D002237:Carbohydrate Dehydrogenases
keywords: iron-sulfur cluster fold; radical SAM enzyme; twitch domain
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