skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: An Iron(IV)–Oxo Intermediate Initiating l-Arginine Oxidation but Not Ethylene Production by the 2-Oxoglutarate-Dependent Oxygenase, Ethylene-Forming Enzyme

Journal Article · · Journal of the American Chemical Society
DOI:https://doi.org/10.1021/jacs.0c10923· OSTI ID:1774768
ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]
  1. Pennsylvania State Univ., University Park, PA (United States)
+ Show Author Affiliations

Ethylene-forming enzyme (EFE) is an ambifunctional iron(II)- and 2-oxoglutarate-dependent (Fe/2OG) oxygenase. In its major (EF) reaction, it converts carbons 1, 2, and 5 of 2OG to CO2 and carbons 3 and 4 to ethylene, a four-electron oxidation drastically different from the simpler decarboxylation of 2OG to succinate mediated by all other Fe/2OG enzymes. EFE also catalyzes a minor reaction, in which the normal decarboxylation is coupled to oxidation of L-arginine (a required activator for the EF pathway), resulting in its conversion to L-glutamate semialdehyde and guanidine. Here we show that, consistent with precedent, the L-Arg-oxidation (RO) pathway proceeds via an iron(IV)-oxo (ferryl) intermediate. Use of 5,5-[2H2]-L-Arg slows decay of the ferryl complex by >16-fold, implying that RO is initiated by hydrogen-atom transfer (HAT) from C5. That this large substrate deuterium kinetic isotope effect has no impact on the EF:RO partition ratio implies that the same ferryl intermediate cannot be on the EF pathway; the pathways must diverge earlier. Consistent with this conclusion, the variant enzyme bearing the Asp191Glu ligand substitution accumulates ~ four times as much of the ferryl complex as the wild-type enzyme and exhibits a ~ 40-fold diminished EF:RO partition ratio. Here, the selective detriment of this nearly conservative substitution to the EF pathway implies that it has unusually stringent stereoelectronic requirements. An active-site, like-charge guanidium pair, which involves the L-Arg substrate/activator and is unique to EFE among four crystallographically characterized L-Arg modifying Fe/2OG oxygenases, may serve to selectively stabilize the transition state leading to the unique EF branch.

Research Organization:
Pennsylvania State Univ., University Park, PA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
Grant/Contract Number:
SC0016255; AC02-06CH11357; AC02-05CH11231
OSTI ID:
1774768
Journal Information:
Journal of the American Chemical Society, Vol. 143, Issue 5; ISSN 0002-7863
Publisher:
American Chemical Society (ACS)Copyright Statement
Country of Publication:
United States
Language:
English

References (47)

Two Distinct Mechanisms for C–C Desaturation by Iron(II)- and 2-(Oxo)glutarate-Dependent Oxygenases: Importance of α-Heteroatom Assistance journal April 2018
Biochemical and Spectroscopic Characterization of the Non-Heme Fe(II)- and 2-Oxoglutarate-Dependent Ethylene-Forming Enzyme from Pseudomonas syringae pv. phaseolicola PK2 journal October 2016
Correction to Biochemical and Spectroscopic Characterization of the Non-Heme Fe(II)- and 2-Oxoglutarate-Dependent Ethylene-Forming Enzyme from Pseudomonas syringae pv. phaseolicola PK2 journal June 2017
Kinetic Dissection of the Catalytic Mechanism of Taurine:α-Ketoglutarate Dioxygenase (TauD) from Escherichia coli † journal June 2005
Electronic Structure Analysis of the Oxygen-Activation Mechanism by Fe II - and α-Ketoglutarate (αKG)-Dependent Dioxygenases journal April 2012
Circular Dichroism and Magnetic Circular Dichroism Spectroscopic Studies of the Non-Heme Ferrous Active Site in Clavaminate Synthase and Its Interaction with α-Ketoglutarate Cosubstrate journal February 1998
Thermodynamics of Iron(II) and Substrate Binding to the Ethylene-Forming Enzyme journal September 2018
Electrostatics-defying interaction between arginine termini as a thermodynamic driving force in protein-protein interaction journal January 2009
Metal ligand substitution and evidence for quinone formation in taurine/α-ketoglutarate dioxygenase journal May 2007
What stabilizes close arginine pairing in proteins? journal January 2013
VioC is a Non-Heme Iron, α-Ketoglutarate-Dependent Oxygenase that Catalyzes the Formation of 3S-Hydroxy-L-Arginine during Viomycin Biosynthesis journal September 2004
Structural basis for the erythro-stereospecificity of the l-arginine oxygenase VioC in viomycin biosynthesis: High-resolution structures of VioC journal June 2009
The First Direct Characterization of a High-Valent Iron Intermediate in the Reaction of an α-Ketoglutarate-Dependent Dioxygenase:  A High-Spin Fe(IV) Complex in Taurine/α-Ketoglutarate Dioxygenase (TauD) from Escherichia coli † journal June 2003
Evidence for Modulation of Oxygen Rebound Rate in Control of Outcome by Iron(II)- and 2-Oxoglutarate-Dependent Oxygenases journal September 2019
Geometric and Electronic Structure/Function Correlations in Non-Heme Iron Enzymes journal January 2000
Mechanistic insights into a non-heme 2-oxoglutarate-dependent ethylene-forming enzyme: selectivity of ethylene-formation versus l -Arg hydroxylation journal January 2019
Comparative sequence analysis and mutagenesis of Ethylene Forming Enzyme (EFE) 2-oxoglutarate/Fe(II)-dependent dioxygenase homologs journal January 2014
Non-Heme Fe(IV)–Oxo Intermediates journal July 2007
Catalytic Mechanisms of Fe(II)- and 2-Oxoglutarate-dependent Oxygenases journal July 2015
The biosynthesis of penicillins and cephalosporins journal January 1988
Changes in Regioselectivity of H Atom Abstraction during the Hydroxylation and Cyclization Reactions Catalyzed by Hyoscyamine 6β-Hydroxylase journal December 2018
Structural Basis for a Kolbe-Type Decarboxylation Catalyzed by a Glycyl Radical Enzyme journal September 2011
Biosynthesis of Streptolidine Involved Two Unexpected Intermediates Produced by a Dihydroxylase and a Cyclase through Unusual Mechanisms journal January 2014
Studies on Deacetoxycephalosporin C Synthase Support a Consensus Mechanism for 2-Oxoglutarate Dependent Oxygenases journal April 2014
Purification and some properties of a novel ethylene-forming enzyme produced by Penicillium digitatum journal May 1989
The 2-His-1-carboxylate facial triad: a versatile platform for dioxygen activation by mononuclear non-heme iron(II) enzymes journal March 2005
Substrate Binding Mode and Molecular Basis of a Specificity Switch in Oxalate Decarboxylase journal March 2016
Substrate positioning controls the partition between halogenation and hydroxylation in the aliphatic halogenase, SyrB2 journal October 2009
Two reactions are simultaneously catalyzed by a single enzyme: The arginine-dependent simultaneous formation of two products, ethylene and succinate, from 2-oxoglutarate by an enzyme from Pseudomonas syringae journal October 1992
The Stabilization Effect of Dielectric Constant and Acidic Amino Acids on Arginine–Arginine (Arg–Arg) Pairings: Database Survey and Computational Studies journal April 2013
Structural and stereoelectronic insights into oxygenase-catalyzed formation of ethylene from 2-oxoglutarate journal April 2017
Oxygen Activation and Radical Transformations in Heme Proteins and Metalloporphyrins journal December 2017
Oxalate Decarboxylase Requires Manganese and Dioxygen for Activity: OVEREXPRESSION AND CHARACTERIZATION OF journal August 2001
Dioxygen activation by nonheme iron enzymes with the 2-His-1-carboxylate facial triad that generate high-valent oxoiron oxidants journal January 2017
Stalking intermediates in oxygen activation by iron enzymes: Motivation and method journal April 2006
Structural studies on 2-oxoglutarate oxygenases and related double-stranded β-helix fold proteins journal April 2006
Direct spectroscopic detection of a C-H-cleaving high-spin Fe(IV) complex in a prolyl-4-hydroxylase journal September 2006
Site-directed mutagenesis of histidine residues in the ethylene-forming enzyme from Pseudomonas syringae journal January 1998
Fe(II)/α-Ketoglutarate-Dependent Hydroxylases and Related Enzymes journal January 2004
Structures and Mechanisms of the Non-Heme Fe(II)- and 2-Oxoglutarate-Dependent Ethylene-Forming Enzyme: Substrate Binding Creates a Twist journal August 2017
Visualizing the Reaction Cycle in an Iron(II)- and 2-(Oxo)-glutarate-Dependent Hydroxylase journal September 2017
The facial triad in the α-ketoglutarate dependent oxygenase FIH: A role for sterics in linking substrate binding to O2 activation journal January 2017
Conversion of (2S)-Arginine to (2S,3R)-Capreomycidine by VioC and VioD from the Viomycin Biosynthetic Pathway of Streptomyces sp. Strain ATCC11861 journal September 2004
Core Assembly Mechanism of Quinocarcin/SF-1739: Bimodular Complex Nonribosomal Peptide Synthetases for Sequential Mannich-type Reactions journal December 2013
The Molecular Origin of Like-Charge Arginine−Arginine Pairing in Water journal July 2009
Dioxygen Activation at Mononuclear Nonheme Iron Active Sites:  Enzymes, Models, and Intermediates journal February 2004
Spectroscopic and Computational Evaluation of the Structure of the High-Spin Fe(IV)-Oxo Intermediates in Taurine: α-Ketoglutarate Dioxygenase from Escherichia coli and Its His99Ala Ligand Variant journal May 2007

Similar Records

α-Amine Desaturation of d-Arginine by the Iron(II)- and 2-(Oxo)glutarate-Dependent l-Arginine 3-Hydroxylase, VioC
Journal Article · Mon Oct 15 00:00:00 EDT 2018 · Biochemistry · OSTI ID:1774768
+3 more
Hybrid radical-polar pathway for excision of ethylene from 2-oxoglutarate by an iron oxygenase
Journal Article · Fri Sep 24 00:00:00 EDT 2021 · Science · OSTI ID:1774768
+3 more
Characterization and structure of the human lysine-2-oxoglutarate reductase domain, a novel therapeutic target for treatment of glutaric aciduria type 1
Journal Article · Wed Sep 21 00:00:00 EDT 2022 · Open Biology · OSTI ID:1774768
+8 more

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
Hydrocarbons
Hydrogen isotopes
Peptides and proteins
Absorption
Chemical reactions