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FEBS Lett. 2017 Feb;591(3):468-478. doi: 10.1002/1873-3468.12554. Epub 2017 Jan 25.

Evidence that oxidative dephosphorylation by the nonheme Fe(II), α-ketoglutarate:UMP oxygenase occurs by stereospecific hydroxylation.

Author information

1
Department of Pharmaceutical Sciences, University of Kentucky, Lexington, KY, USA.
2
Department of Pharmaceutical Sciences, University of Wisconsin-Madison, WI, USA.
3
Nucleosides and Phosphorylated Effectors Team, IBMM, UMR5247 CNRS University Montpellier, France.
4
Biologics Technology Research Laboratories, R&D Division, Daiichi Sankyo Co., Ltd., Gunma, Japan.

Abstract

LipL and Cpr19 are nonheme, mononuclear Fe(II)-dependent, α-ketoglutarate (αKG):UMP oxygenases that catalyze the formation of CO2 , succinate, phosphate, and uridine-5'-aldehyde, the last of which is a biosynthetic precursor for several nucleoside antibiotics that inhibit bacterial translocase I (MraY). To better understand the chemistry underlying this unusual oxidative dephosphorylation and establish a mechanistic framework for LipL and Cpr19, we report herein the synthesis of two biochemical probes-[1',3',4',5',5'-2 H]UMP and the phosphonate derivative of UMP-and their activity with both enzymes. The results are consistent with a reaction coordinate that proceeds through the loss of one 2 H atom of [1',3',4',5',5'-2 H]UMP and stereospecific hydroxylation geminal to the phosphoester to form a cryptic intermediate, (5'R)-5'-hydroxy-UMP. Thus, these enzyme catalysts can additionally be assigned as UMP hydroxylase-phospholyases.

KEYWORDS:

antibiotic; biosynthesis; nonheme iron; nucleoside; oxygenase; translocase I

PMID:
28074470
PMCID:
PMC5380139
DOI:
10.1002/1873-3468.12554
[Indexed for MEDLINE]
Free PMC Article

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