Biochemical and Spectroscopic Studies of Epoxyqueuosine Reductase: A Novel Iron-Sulfur Cluster- and Cobalamin-Containing Protein Involved in the Biosynthesis of Queuosine

Zachary D. Miles, William K. Myers, William M. Kincannon, R. David Britt, Vahe Bandarian

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Queuosine is a hypermodified nucleoside present in the wobble position of tRNAs with a 5′-GUN-3′ sequence in their anticodon (His, Asp, Asn, and Tyr). The 7-deazapurine core of the base is synthesized de novo in prokaryotes from guanosine 5′-triphosphate in a series of eight sequential enzymatic transformations, the final three occurring on tRNA. Epoxyqueuosine reductase (QueG) catalyzes the final step in the pathway, which entails the two-electron reduction of epoxyqueuosine to form queuosine. Biochemical analyses reveal that this enzyme requires cobalamin and two [4Fe-4S] clusters for catalysis. Spectroscopic studies show that the cobalamin appears to bind in a base-off conformation, whereby the dimethylbenzimidazole moiety of the cofactor is removed from the coordination sphere of the cobalt but not replaced by an imidazole side chain, which is a hallmark of many cobalamin-dependent enzymes. The bioinformatically identified residues are shown to have a role in modulating the primary coordination sphere of cobalamin. These studies provide the first demonstration of the cofactor requirements for QueG.

Original languageEnglish (US)
Pages (from-to)4927-4935
Number of pages9
JournalBiochemistry
Volume54
Issue number31
DOIs
StatePublished - Aug 11 2015

ASJC Scopus subject areas

  • Biochemistry

Fingerprint Dive into the research topics of 'Biochemical and Spectroscopic Studies of Epoxyqueuosine Reductase: A Novel Iron-Sulfur Cluster- and Cobalamin-Containing Protein Involved in the Biosynthesis of Queuosine'. Together they form a unique fingerprint.

  • Cite this