Kinetics of Nitrite Reduction and Peroxynitrite Formation by Ferrous Heme in Human Cystathionine β-Synthase

Carballal, Sebastián; Cuevasanta, Ernesto; Yadav, Pramod Kumar; Gherasim, Carmen; Ballou, David P.; Álvarez, Beatriz; Banerjee, Ruma

doi:10.1074/jbc.m116.718734

Cited by 22 publications

(21 citation statements)

References 77 publications

(65 reference statements)

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“…The N-terminal domain binds the heme cofactor, which seems to function as a redox sensor (13) that inhibits CBS activity by means of binding of CO or NO (14)(15)(16)(17) or during reduction of nitrite (18,19). Nitration of tryptophan residues in CBS results in alterations of the heme pocket, which lead to the loss of cysteinate coordination by the ligand Cys52 and concomitant inactivation of CBS (20).…”

Section: Introductionmentioning

confidence: 99%

Allosteric control of human cystathionine β-synthase activity by a redox active disulfide bond

Niu¹,

Wang²,

Qian

et al. 2018

Journal of Biological Chemistry

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Cystathionine β-synthase (CBS) is the central enzyme in the trans-sulfuration pathway that converts homocysteine to cysteine. It is also one of the three major enzymes involved in the biogenesis of H 2 S. CBS is a complex protein with a modular three-domain architecture, the central domain of which contains a C 272 XXC 275 motif whose function has yet to be determined. In the present study, we demonstrated that the CXXC motif exists in oxidized and reduced states in the recombinant enzyme by mass spectroscopic analysis and a thiol labeling assay. The activity of reduced CBS is ~2-to 3-fold greater than that of the oxidized enzyme, and substitution of either cysteine in CXXC motif leads to a loss of redox sensitivity. The Cys 272 -Cys 275 disulfide bond in CBS has a midpoint potential of -314 mV at pH 7.4. Additionally, the CXXC motif also exists in oxidized and reduced states in human embryonic kidney 293 (HEK293) cells under oxidative and reductive conditions, and stressing these cells with dithiothreitol (DTT) results in more reduced enzyme and a concomitant increase in H 2 S production in live HEK293 cells as determined using a H 2 S fluorescent probe. By contrast, incubation of cells with aminooxyacetic acid (AOAA), an inhibitor of CBS and cystathionine γ-lyase (CSE), eliminates the increase of H 2 S production after the cells were exposed to DTT. These findings indicate that CBS is post-translationally regulated by a redox-active disulfide bond in the CXXC motif. The results als o demonstrate that CBS-derived H 2 S production is increased in cells under reductive stress conditions.

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Section: Introductionmentioning

confidence: 99%

Allosteric control of human cystathionine β-synthase activity by a redox active disulfide bond

Niu¹,

Wang²,

Qian

et al. 2018

Journal of Biological Chemistry

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“…A close homolog to LmPP with respect to heme ligation is cystathionine-β-synthase (CBS), but unlike LmPP, CBS has been extensively studied and yet the function of the heme remains unknown. However, like CBS [41], the heme iron in LmPP can be reduced with dithionite, enabling formation of the ferrous-carbon monoxide (CO) complex. The spectral features of the CO complex are characteristic of His-ligation (Figure S6).…”

Section: Discussionmentioning

confidence: 99%

Crystal structure and functional analysis of Leishmania major pseudoperoxidase

2017

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Leishmania major pseudoperoxidase (LmPP) is a recently discovered heme protein expressed by the human pathogen. Previous in vivo and in vitro studies suggest that LmPP is a crucial element of the pathogen’s defense mechanism against the reactive nitrogen species peroxynitrite produced during the host immune response. In order to shed light on the potential mechanism of peroxynitrite detoxification, we have determined the 1.76-Å X-ray crystal structure of LmPP, revealing a striking degree of homology with heme peroxidases. The most outstanding structural feature is a Cys/His heme coordination, which corroborates previous spectroscopic and mutagenesis studies. We also used a combination of stopped-flow and electron paramagnetic spectroscopies that together suggest that peroxynitrite is not a substrate for LmPP catalysis, leaving the function of LmPP an open question.

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“…The reaction of oxidized CBS with peroxynitrite has been shown to result in enzyme inactivation with an IC 50 of 150 μ M, occurring according to a bimolecular rate constant of 2.4–5 × 10 4 M −1 ·s −1 and leading to nitration of Trp 208 , Trp 43 , and Tyr 223 , as well as damaging the heme moiety [ 215 ]. Intriguingly, a recent report demonstrated that the reaction between ferrous-NO CBS and O 2 , previously shown to revert the heme to its ferric state, yields peroxynitrite [ 201 ] ( Figure 13 ). Moreover, the reaction of reduced CBS with nitrite may itself constitute a source of NO [ 191 , 201 ], which further adds to the complex heme-mediated regulation of H 2 S synthesis by CBS ( Figure 13 ).…”

Section: Interplay Between Gasotransmittersmentioning

confidence: 96%

“…Intriguingly, a recent report demonstrated that the reaction between ferrous-NO CBS and O 2 , previously shown to revert the heme to its ferric state, yields peroxynitrite [ 201 ] ( Figure 13 ). Moreover, the reaction of reduced CBS with nitrite may itself constitute a source of NO [ 191 , 201 ], which further adds to the complex heme-mediated regulation of H 2 S synthesis by CBS ( Figure 13 ).…”

Section: Interplay Between Gasotransmittersmentioning

confidence: 96%

Hydrogen Sulfide Biochemistry and Interplay with Other Gaseous Mediators in Mammalian Physiology

Giuffrè

Vicente

2018

Oxidative Medicine and Cellular Longevity

107

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Hydrogen sulfide (H2S) has emerged as a relevant signaling molecule in physiology, taking its seat as a bona fide gasotransmitter akin to nitric oxide (NO) and carbon monoxide (CO). After being merely regarded as a toxic poisonous molecule, it is now recognized that mammalian cells are equipped with sophisticated enzymatic systems for H2S production and breakdown. The signaling role of H2S is mainly related to its ability to modify different protein targets, particularly by promoting persulfidation of protein cysteine residues and by interacting with metal centers, mostly hemes. H2S has been shown to regulate a myriad of cellular processes with multiple physiological consequences. As such, dysfunctional H2S metabolism is increasingly implicated in different pathologies, from cardiovascular and neurodegenerative diseases to cancer. As a highly diffusible reactive species, the intra- and extracellular levels of H2S have to be kept under tight control and, accordingly, regulation of H2S metabolism occurs at different levels. Interestingly, even though H2S, NO, and CO have similar modes of action and parallel regulatory targets or precisely because of that, there is increasing evidence of a crosstalk between the three gasotransmitters. Herein are reviewed the biochemistry, metabolism, and signaling function of hydrogen sulfide, as well as its interplay with the other gasotransmitters, NO and CO.

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Kinetics of Nitrite Reduction and Peroxynitrite Formation by Ferrous Heme in Human Cystathionine β-Synthase

Cited by 22 publications

References 77 publications

Allosteric control of human cystathionine β-synthase activity by a redox active disulfide bond

Allosteric control of human cystathionine β-synthase activity by a redox active disulfide bond

Crystal structure and functional analysis of Leishmania major pseudoperoxidase

Hydrogen Sulfide Biochemistry and Interplay with Other Gaseous Mediators in Mammalian Physiology

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