Human Cystathionine γ-Lyase Is Inhibited by <i>s</i>-Nitrosation: A New Crosstalk Mechanism between NO and H<sub>2</sub>S
The ‘gasotransmitters’ hydrogen sulfide (H<sub>2</sub>S), nitric oxide (NO), and carbon monoxide (CO) act as second messengers in human physiology, mediating signal transduction via interaction with or chemical modification of protein targets, thereby regulating processes such as neurotr...
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MDPI AG
2021-08-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/10/9/1391 |
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| author | Dalila G. F. Fernandes João Nunes Catarina S. Tomé Karim Zuhra João M. F. Costa Alexandra M. M. Antunes Alessandro Giuffrè João B. Vicente |
| author_facet | Dalila G. F. Fernandes João Nunes Catarina S. Tomé Karim Zuhra João M. F. Costa Alexandra M. M. Antunes Alessandro Giuffrè João B. Vicente |
| author_sort | Dalila G. F. Fernandes |
| collection | DOAJ |
| description | The ‘gasotransmitters’ hydrogen sulfide (H<sub>2</sub>S), nitric oxide (NO), and carbon monoxide (CO) act as second messengers in human physiology, mediating signal transduction via interaction with or chemical modification of protein targets, thereby regulating processes such as neurotransmission, blood flow, immunomodulation, or energy metabolism. Due to their broad reactivity and potential toxicity, the biosynthesis and breakdown of H<sub>2</sub>S, NO, and CO are tightly regulated. Growing evidence highlights the active role of gasotransmitters in their mutual cross-regulation. In human physiology, the transsulfuration enzymes cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE) are prominent H<sub>2</sub>S enzymatic sources. While CBS is known to be inhibited by NO and CO, little is known about CSE regulation by gasotransmitters. Herein, we investigated the effect of <i>s</i>-nitrosation on CSE catalytic activity. H<sub>2</sub>S production by recombinant human CSE was found to be inhibited by the physiological nitrosating agent <i>s</i>-nitrosoglutathione (GSNO), while reduced glutathione had no effect. GSNO-induced inhibition was partially reverted by ascorbate and accompanied by the disappearance of one solvent accessible protein thiol. By combining differential derivatization procedures and mass spectrometry-based analysis with functional assays, seven out of the ten protein cysteine residues, namely Cys84, Cys109, Cys137, Cys172, Cys229, Cys307, and Cys310, were identified as targets of <i>s</i>-nitrosation. By generating conservative Cys-to-Ser variants of the identified <i>s</i>-nitrosated cysteines, Cys137 was identified as most significantly contributing to the GSNO-mediated CSE inhibition. These results highlight a new mechanism of crosstalk between gasotransmitters. |
| first_indexed | 2024-03-10T07:57:26Z |
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| id | doaj.art-c4183674574e4809bfd528d33eee1c84 |
| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-10T07:57:26Z |
| publishDate | 2021-08-01 |
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| series | Antioxidants |
| spelling | doaj.art-c4183674574e4809bfd528d33eee1c842023-11-22T11:48:02ZengMDPI AGAntioxidants2076-39212021-08-01109139110.3390/antiox10091391Human Cystathionine γ-Lyase Is Inhibited by <i>s</i>-Nitrosation: A New Crosstalk Mechanism between NO and H<sub>2</sub>SDalila G. F. Fernandes0João Nunes1Catarina S. Tomé2Karim Zuhra3João M. F. Costa4Alexandra M. M. Antunes5Alessandro Giuffrè6João B. Vicente7Instituto de Tecnologia Química e Biológica António Xavier (ITQB NOVA), 2780-157 Oeiras, PortugalCentro de Química Estrutural, Instituto Superior Técnico, ULisboa, 1049-001 Lisboa, PortugalInstituto de Tecnologia Química e Biológica António Xavier (ITQB NOVA), 2780-157 Oeiras, PortugalCNR Institute of Molecular Biology and Pathology, I-00185 Rome, ItalyInstituto de Tecnologia Química e Biológica António Xavier (ITQB NOVA), 2780-157 Oeiras, PortugalCentro de Química Estrutural, Instituto Superior Técnico, ULisboa, 1049-001 Lisboa, PortugalCNR Institute of Molecular Biology and Pathology, I-00185 Rome, ItalyInstituto de Tecnologia Química e Biológica António Xavier (ITQB NOVA), 2780-157 Oeiras, PortugalThe ‘gasotransmitters’ hydrogen sulfide (H<sub>2</sub>S), nitric oxide (NO), and carbon monoxide (CO) act as second messengers in human physiology, mediating signal transduction via interaction with or chemical modification of protein targets, thereby regulating processes such as neurotransmission, blood flow, immunomodulation, or energy metabolism. Due to their broad reactivity and potential toxicity, the biosynthesis and breakdown of H<sub>2</sub>S, NO, and CO are tightly regulated. Growing evidence highlights the active role of gasotransmitters in their mutual cross-regulation. In human physiology, the transsulfuration enzymes cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE) are prominent H<sub>2</sub>S enzymatic sources. While CBS is known to be inhibited by NO and CO, little is known about CSE regulation by gasotransmitters. Herein, we investigated the effect of <i>s</i>-nitrosation on CSE catalytic activity. H<sub>2</sub>S production by recombinant human CSE was found to be inhibited by the physiological nitrosating agent <i>s</i>-nitrosoglutathione (GSNO), while reduced glutathione had no effect. GSNO-induced inhibition was partially reverted by ascorbate and accompanied by the disappearance of one solvent accessible protein thiol. By combining differential derivatization procedures and mass spectrometry-based analysis with functional assays, seven out of the ten protein cysteine residues, namely Cys84, Cys109, Cys137, Cys172, Cys229, Cys307, and Cys310, were identified as targets of <i>s</i>-nitrosation. By generating conservative Cys-to-Ser variants of the identified <i>s</i>-nitrosated cysteines, Cys137 was identified as most significantly contributing to the GSNO-mediated CSE inhibition. These results highlight a new mechanism of crosstalk between gasotransmitters.https://www.mdpi.com/2076-3921/10/9/1391hydrogen sulfide<i>s</i>-nitrosoglutathionegasotransmitterscystathionine γ-lyasesignalingcrosstalk |
| spellingShingle | Dalila G. F. Fernandes João Nunes Catarina S. Tomé Karim Zuhra João M. F. Costa Alexandra M. M. Antunes Alessandro Giuffrè João B. Vicente Human Cystathionine γ-Lyase Is Inhibited by <i>s</i>-Nitrosation: A New Crosstalk Mechanism between NO and H<sub>2</sub>S Antioxidants hydrogen sulfide <i>s</i>-nitrosoglutathione gasotransmitters cystathionine γ-lyase signaling crosstalk |
| title | Human Cystathionine γ-Lyase Is Inhibited by <i>s</i>-Nitrosation: A New Crosstalk Mechanism between NO and H<sub>2</sub>S |
| title_full | Human Cystathionine γ-Lyase Is Inhibited by <i>s</i>-Nitrosation: A New Crosstalk Mechanism between NO and H<sub>2</sub>S |
| title_fullStr | Human Cystathionine γ-Lyase Is Inhibited by <i>s</i>-Nitrosation: A New Crosstalk Mechanism between NO and H<sub>2</sub>S |
| title_full_unstemmed | Human Cystathionine γ-Lyase Is Inhibited by <i>s</i>-Nitrosation: A New Crosstalk Mechanism between NO and H<sub>2</sub>S |
| title_short | Human Cystathionine γ-Lyase Is Inhibited by <i>s</i>-Nitrosation: A New Crosstalk Mechanism between NO and H<sub>2</sub>S |
| title_sort | human cystathionine γ lyase is inhibited by i s i nitrosation a new crosstalk mechanism between no and h sub 2 sub s |
| topic | hydrogen sulfide <i>s</i>-nitrosoglutathione gasotransmitters cystathionine γ-lyase signaling crosstalk |
| url | https://www.mdpi.com/2076-3921/10/9/1391 |
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