Mutation in the flavin mononucleotide domain modulates magnetic circular dichroism spectra of the iNOS ferric cyano complex in a substrate-specific manner

Joseph Sempombe, Mary Grace I. Galinato, Bradley O. Elmore, Weihong Fan, J. Guy Guillemette, Nicolai Lehnert, Martin L. Kirk, Changjian Feng

Research output: Contribution to journalArticle

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Abstract

We have obtained low-temperature magnetic circular dichroism (MCD) spectra for ferric cyano complexes of the wild type and E546N mutant of a human inducible nitric oxide synthase (iNOS) oxygenase/flavin mononucleotide (oxyFMN) construct. The mutation at the FMN domain has previously been shown to modulate the MCD spectra of the l-arginine-bound ferric iNOS heme (Sempombe, J.; et al. J. Am. Chem. Soc.2009, 131, 6940-6941). The addition of l-arginine to the wild-type protein causes notable changes in the CN --adduct MCD spectrum, while the E546N mutant spectrum is not perturbed. Moreover, the MCD spectral perturbation observed with l-arginine is absent in the CN - complexes incubated with N-hydroxy-l-arginine, which is the substrate for the second step of NOS catalysis. These results indicate that interdomain FMN-heme interactions exert a long-range effect on key heme axial ligand-substrate interactions that determine substrate oxidation pathways of NOS.

Original languageEnglish (US)
Pages (from-to)6859-6861
Number of pages3
JournalInorganic chemistry
Volume50
Issue number15
DOIs
StatePublished - Aug 1 2011

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Flavin Mononucleotide
Magnetic domains
Dichroism
Nitric Oxide Synthase Type II
nitric oxide
magnetic domains
mutations
dichroism
Arginine
Heme
Substrates
Oxygenases
Catalysis
adducts
catalysis
interactions
Ligands
proteins
perturbation
Oxidation

All Science Journal Classification (ASJC) codes

  • Physical and Theoretical Chemistry
  • Inorganic Chemistry

Cite this

Sempombe, Joseph ; Galinato, Mary Grace I. ; Elmore, Bradley O. ; Fan, Weihong ; Guillemette, J. Guy ; Lehnert, Nicolai ; Kirk, Martin L. ; Feng, Changjian. / Mutation in the flavin mononucleotide domain modulates magnetic circular dichroism spectra of the iNOS ferric cyano complex in a substrate-specific manner. In: Inorganic chemistry. 2011 ; Vol. 50, No. 15. pp. 6859-6861.
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abstract = "We have obtained low-temperature magnetic circular dichroism (MCD) spectra for ferric cyano complexes of the wild type and E546N mutant of a human inducible nitric oxide synthase (iNOS) oxygenase/flavin mononucleotide (oxyFMN) construct. The mutation at the FMN domain has previously been shown to modulate the MCD spectra of the l-arginine-bound ferric iNOS heme (Sempombe, J.; et al. J. Am. Chem. Soc.2009, 131, 6940-6941). The addition of l-arginine to the wild-type protein causes notable changes in the CN --adduct MCD spectrum, while the E546N mutant spectrum is not perturbed. Moreover, the MCD spectral perturbation observed with l-arginine is absent in the CN - complexes incubated with N-hydroxy-l-arginine, which is the substrate for the second step of NOS catalysis. These results indicate that interdomain FMN-heme interactions exert a long-range effect on key heme axial ligand-substrate interactions that determine substrate oxidation pathways of NOS.",
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Mutation in the flavin mononucleotide domain modulates magnetic circular dichroism spectra of the iNOS ferric cyano complex in a substrate-specific manner. / Sempombe, Joseph; Galinato, Mary Grace I.; Elmore, Bradley O.; Fan, Weihong; Guillemette, J. Guy; Lehnert, Nicolai; Kirk, Martin L.; Feng, Changjian.

In: Inorganic chemistry, Vol. 50, No. 15, 01.08.2011, p. 6859-6861.

Research output: Contribution to journalArticle

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AU - Sempombe, Joseph

AU - Galinato, Mary Grace I.

AU - Elmore, Bradley O.

AU - Fan, Weihong

AU - Guillemette, J. Guy

AU - Lehnert, Nicolai

AU - Kirk, Martin L.

AU - Feng, Changjian

PY - 2011/8/1

Y1 - 2011/8/1

N2 - We have obtained low-temperature magnetic circular dichroism (MCD) spectra for ferric cyano complexes of the wild type and E546N mutant of a human inducible nitric oxide synthase (iNOS) oxygenase/flavin mononucleotide (oxyFMN) construct. The mutation at the FMN domain has previously been shown to modulate the MCD spectra of the l-arginine-bound ferric iNOS heme (Sempombe, J.; et al. J. Am. Chem. Soc.2009, 131, 6940-6941). The addition of l-arginine to the wild-type protein causes notable changes in the CN --adduct MCD spectrum, while the E546N mutant spectrum is not perturbed. Moreover, the MCD spectral perturbation observed with l-arginine is absent in the CN - complexes incubated with N-hydroxy-l-arginine, which is the substrate for the second step of NOS catalysis. These results indicate that interdomain FMN-heme interactions exert a long-range effect on key heme axial ligand-substrate interactions that determine substrate oxidation pathways of NOS.

AB - We have obtained low-temperature magnetic circular dichroism (MCD) spectra for ferric cyano complexes of the wild type and E546N mutant of a human inducible nitric oxide synthase (iNOS) oxygenase/flavin mononucleotide (oxyFMN) construct. The mutation at the FMN domain has previously been shown to modulate the MCD spectra of the l-arginine-bound ferric iNOS heme (Sempombe, J.; et al. J. Am. Chem. Soc.2009, 131, 6940-6941). The addition of l-arginine to the wild-type protein causes notable changes in the CN --adduct MCD spectrum, while the E546N mutant spectrum is not perturbed. Moreover, the MCD spectral perturbation observed with l-arginine is absent in the CN - complexes incubated with N-hydroxy-l-arginine, which is the substrate for the second step of NOS catalysis. These results indicate that interdomain FMN-heme interactions exert a long-range effect on key heme axial ligand-substrate interactions that determine substrate oxidation pathways of NOS.

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