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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
 [1];  [2];  [2];  [2];  [2];  [2];  [2]
  1. Pennsylvania State Univ., University Park, PA (United States); The Pennsylvania State University
  2. 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:
AC02-05CH11231; AC02-06CH11357; SC0016255
OSTI ID:
1774768
Alternate ID(s):
OSTI ID: 1771382
OSTI ID: 1772778
Journal Information:
Journal of the American Chemical Society, Journal Name: Journal of the American Chemical Society Journal Issue: 5 Vol. 143; ISSN 0002-7863
Publisher:
American Chemical Society (ACS)Copyright Statement
Country of Publication:
United States
Language:
English

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