Substrate Promotes Productive Gas Binding in the α-Ketoglutarate-Dependent Oxygenase FIH

Taabazuing, Cornelius Y.; Fermann, Justin; Garman, Scott; Knapp, Michael J.

doi:10.1021/acs.biochem.5b01003

Title: Substrate Promotes Productive Gas Binding in the α-Ketoglutarate-Dependent Oxygenase FIH

Full Record
Other Related Research

Abstract

The Fe²⁺/α-ketoglutarate (αKG)-dependent oxygenases use molecular oxygen to conduct a wide variety of reactions with important biological implications, such as DNA base excision repair, histone demethylation, and the cellular hypoxia response. These enzymes follow a sequential mechanism in which O₂ binds and reacts after the primary substrate binds, making those structural factors that promote productive O₂ binding central to their chemistry. A large challenge in this field is to identify strategies that engender productive turnover. Factor inhibiting HIF (FIH) is a Fe²⁺/αKG-dependent oxygenase that forms part of the O₂ sensing machinery in human cells by hydroxylating the C-terminal transactivation domain (CTAD) found within the HIF-1α protein. In this work, the structure of FIH was determined with the O₂ analogue NO bound to Fe, offering the first direct insight into the gas binding geometry in this enzyme. Through a combination of density functional theory calculations, {FeNO}⁷ electron paramagnetic resonance spectroscopy, and ultraviolet–visible absorption spectroscopy, we demonstrate that CTAD binding stimulates O₂ reactivity by altering the orientation of the bound gas molecule. Although unliganded FIH binds NO with moderate affinity, the bound gas can adopt either of two orientations with similar stability; upon CTAD binding, NO adopts a single preferred orientation thatmore »« less

Authors:

Taabazuing, Cornelius Y. ^[1]; Fermann, Justin ^[1]; Garman, Scott ^[1]; Knapp, Michael J. ^[1]

Univ. of Massachusetts, Amherst, MA (United States)

Publication Date:: Tue Jan 05 00:00:00 EST 2016

Research Org.:: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Org.:: USDOE Office of Science (SC); National Institutes of Health (NIH); National Science Foundation (NSF); National Institute of General Medical Sciences (NIGMS)

OSTI Identifier:: 1236263

Grant/Contract Number:: AC02-06CH11357; CHE-0443180 NSF-CRIF; P41 GM103403

Resource Type:: Accepted Manuscript

Journal Name:: Biochemistry

Additional Journal Information:: Journal Volume: 55; Journal Issue: 2; Journal ID: ISSN 0006-2960

Publisher:: American Chemical Society (ACS)

Country of Publication:: United States

Language:: ENGLISH

Subject:: 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Peptides and proteins; Surface interactions; Organic reactions; Chemical structure

Citation Formats


                    Taabazuing, Cornelius Y., Fermann, Justin, Garman, Scott, and Knapp, Michael J. Substrate Promotes Productive Gas Binding in the α-Ketoglutarate-Dependent Oxygenase FIH.  United States: N. p., 2016. 
Web.  doi:10.1021/acs.biochem.5b01003.

Copy to clipboard


                    Taabazuing, Cornelius Y., Fermann, Justin, Garman, Scott, & Knapp, Michael J. Substrate Promotes Productive Gas Binding in the α-Ketoglutarate-Dependent Oxygenase FIH.  United States.  https://doi.org/10.1021/acs.biochem.5b01003

Copy to clipboard


                    Taabazuing, Cornelius Y., Fermann, Justin, Garman, Scott, and Knapp, Michael J. Tue .  
"Substrate Promotes Productive Gas Binding in the α-Ketoglutarate-Dependent Oxygenase FIH".  United States.  https://doi.org/10.1021/acs.biochem.5b01003.  https://www.osti.gov/servlets/purl/1236263.

Copy to clipboard


                    
@article{osti_1236263,

  title        = {Substrate Promotes Productive Gas Binding in the α-Ketoglutarate-Dependent Oxygenase FIH},

  author       = {Taabazuing, Cornelius Y. and Fermann, Justin and Garman, Scott and Knapp, Michael J.},

  abstractNote = {The Fe2+/α-ketoglutarate (αKG)-dependent oxygenases use molecular oxygen to conduct a wide variety of reactions with important biological implications, such as DNA base excision repair, histone demethylation, and the cellular hypoxia response. These enzymes follow a sequential mechanism in which O2 binds and reacts after the primary substrate binds, making those structural factors that promote productive O2 binding central to their chemistry. A large challenge in this field is to identify strategies that engender productive turnover. Factor inhibiting HIF (FIH) is a Fe2+/αKG-dependent oxygenase that forms part of the O2 sensing machinery in human cells by hydroxylating the C-terminal transactivation domain (CTAD) found within the HIF-1α protein. In this work, the structure of FIH was determined with the O2 analogue NO bound to Fe, offering the first direct insight into the gas binding geometry in this enzyme. Through a combination of density functional theory calculations, {FeNO}7 electron paramagnetic resonance spectroscopy, and ultraviolet–visible absorption spectroscopy, we demonstrate that CTAD binding stimulates O2 reactivity by altering the orientation of the bound gas molecule. Although unliganded FIH binds NO with moderate affinity, the bound gas can adopt either of two orientations with similar stability; upon CTAD binding, NO adopts a single preferred orientation that is appropriate for supporting oxidative decarboxylation. Combined with other studies of related enzymes, our data suggest that substrate-induced reorientation of bound O2 is the mechanism utilized by the αKG oxygenases to tightly couple O2 activation to substrate hydroxylation.},

  doi          = {10.1021/acs.biochem.5b01003},

  journal      = {Biochemistry},

  number       = 2,

  volume       = 55,

  place        = {United States},

  year         = {Tue Jan 05 00:00:00 EST 2016},

  month        = {Tue Jan 05 00:00:00 EST 2016}

}

Copy to clipboard

Journal Article:

Free Publicly Available Full Text

Accepted Manuscript (DOE)

Publisher's Version of Record

https://doi.org/10.1021/acs.biochem.5b01003

Other availability

Search WorldCat to find libraries that may hold this journal

Citation Metrics:

Cited by: 10 works

Citation information provided by
Web of Science

Save / Share:

Export Metadata

Save to My Library

Similar Records in DOE PAGES and OSTI.GOV collections:

Evaluation of a concerted vs. sequential oxygen activation mechanism in α-ketoglutarate–dependent nonheme ferrous enzymes

Journal Article Goudarzi, Serra ; Iyer, Shyam R. ; Babicz, Jr., Jeffrey T. ; ... - Proceedings of the National Academy of Sciences of the United States of America

Determining the requirements for efficient oxygen (O₂) activation is key to understanding how enzymes maintain efficacy and mitigate unproductive, often detrimental reactivity. For the α-ketoglutarate (αKG)–dependent nonheme iron enzymes, both a concerted mechanism (both cofactor and substrate binding prior to reaction with O₂) and a sequential mechanism (cofactor binding and reaction with O₂precede substrate binding) have been proposed. Deacetoxycephalosporin C synthase (DAOCS) is an αKG-dependent nonheme iron enzyme for which both of these mechanisms have been invoked to generate an intermediate that catalyzes oxidative ring expansion of penicillin substrates in cephalosporin biosynthesis. Interestingly, spectroscopy shows that, in contrast to othermore »« less
Cited by 12
https://doi.org/10.1073/pnas.1922484117

Full Text Available
Ferritin heavy chain controls the HIF-driven hypoxic response by activating the asparaginyl hydroxylase FIH

Journal Article Jin, Peng ; Kang, Jengmin ; Ischemic/Hypoxic Disease Institute and Cancer Research Institute, Seoul National University College of Medicine, Seoul, South ; ... - Biochemical and Biophysical Research Communications

Highlights: • Ferritin Heavy Chain (FTH1) directly interacts with Factor Inhibiting HIF (FIH). • FTH1 facilitates the FIH-mediated Asn803 hydroxylation in HIF-1α. • FTH1 prevents the recruitment of p300 to HIF-1α through the Asn803 hydroxylation. • FTH1 represses the HIF-driven gene expression under either normoxic or hypoxic conditions. Hypoxia-inducible factor 1 (HIF-1) is a key player in cellular response to hypoxia. The stability and transcriptional activity of this protein are oxygen-dependently regulated by the prolyl hydroxylases PHD1-3 and the asparaginyl hydroxylase FIH. Recently, ferritin heavy chain (FTH1) has been characterized to reinforce the HIF-1 signaling pathway in an indirect waymore »« less
https://doi.org/10.1016/J.BBRC.2018.03.173
Coordination Changes And Auto-Hydroxylation of FIH-1: Uncoupled O(2)-Activa in a Human Hypoxia Sensor

Journal Article Chen, Y -H ; Comeaux, L M ; Herbst, R W ; ... - J. Inorg. Biochem 102:2120,2008

Hypoxia sensing is the generic term for pO2-sensing in humans and other higher organisms. These cellular responses to pO2 are largely controlled by enzymes that belong to the Fe(II) alpha-ketoglutarate (alphaKG) dependent dioxygenase superfamily, including the human enzyme called the factor inhibiting HIF (FIH-1), which couples O2-activation to the hydroxylation of the hypoxia inducible factor alpha (HIFalpha). Uncoupled O2-activation by human FIH-1 was studied by exposing the resting form of FIH-1 (alphaKG + Fe)FIH-1, to air in the absence of HIFalpha. Uncoupling lead to two distinct enzyme oxidations, one a purple chromophore (lambda(max) = 583 nm) arising from enzyme auto-hydroxylationmore »« less
Structure and assembly of the diiron cofactor in the heme-oxygenase–like domain of the N-nitrosourea–producing enzyme SznF

Journal Article McBride, Molly J. ; Pope, Sarah R. ; Hu, Kai ; ... - Proceedings of the National Academy of Sciences of the United States of America

In biosynthesis of the pancreatic cancer drug streptozotocin, the tridomain nonheme-iron oxygenase SznF hydroxylates N^δ and N^ω' of N^ω-methyl-L-arginine before oxidatively rearranging the triply modified guanidine to the N-methyl-N-nitrosourea pharmacophore. A previously published structure visualized the monoiron cofactor in the enzyme’s C-terminal cupin domain, which promotes the final rearrangement, but exhibited disorder and minimal metal occupancy in the site of the proposed diiron cofactor in the N-hydroxylating heme-oxygenase–like (HO-like) central domain. We leveraged our recent observation that the N-oxygenating µ-peroxodiiron(III/III) intermediate can form in the HO-like domain after the apo protein self-assembles its diiron(II/II) cofactor to solve structures of SznFmore »« less
https://doi.org/10.1073/pnas.2015931118

Full Text Available
Structural Investigations of the Ferredoxin and Terminal Oxygenase Components of the biphenyl 2,3-dioxygenase from Sphingobium yanoikuyae B1

Journal Article Ferraro, D ; Brown, E ; Yu, C ; ... - BMC Structural Biology

The initial step involved in oxidative hydroxylation of monoaromatic and polyaromatic compounds by the microorganism Sphingobium yanoikuyae strain B1 (B1), previously known as Sphingomonas yanoikuyae strain B1 and Beijerinckia sp. strain B1, is performed by a set of multiple terminal Rieske non-heme iron oxygenases. These enzymes share a single electron donor system consisting of a reductase and a ferredoxin (BPDO-F{sub B1}). One of the terminal Rieske oxygenases, biphenyl 2,3-dioxygenase (BPDO-O{sub B1}), is responsible for B1's ability to dihydroxylate large aromatic compounds, such as chrysene and benzo(a)pyrene. Results: In this study, crystal structures of BPDO-O{sub B1} in both native and biphenylmore »« less
https://doi.org/10.1186/1472-6807-7-10

Similar Records