Biochemical Characterization of the Copper Nitrite Reductase from <i>Neisseria gonorrhoeae</i>
The copper-containing nitrite reductase from <i>Neisseria gonorrhoeae</i> has been shown to play a critical role in the infection mechanism of this microorganism by producing NO and abolishing epithelial exfoliation. This enzyme is a trimer with a type 1 copper center per subunit and a t...
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MDPI AG
2023-08-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/13/8/1215 |
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| author | Daniela S. Barreiro Ricardo N. S. Oliveira Sofia R. Pauleta |
| author_facet | Daniela S. Barreiro Ricardo N. S. Oliveira Sofia R. Pauleta |
| author_sort | Daniela S. Barreiro |
| collection | DOAJ |
| description | The copper-containing nitrite reductase from <i>Neisseria gonorrhoeae</i> has been shown to play a critical role in the infection mechanism of this microorganism by producing NO and abolishing epithelial exfoliation. This enzyme is a trimer with a type 1 copper center per subunit and a type 2 copper center in the subunits interface, with the latter being the catalytic site. The two centers were characterized for the first time by EPR and CD spectroscopy, showing that the type 1 copper center has a high rhombicity due to its lower symmetry and more tetragonal structure, while the type 2 copper center has the usual properties, but with a smaller hyperfine coupling constant (A<sub>//</sub> = 10.5 mT). The thermostability of the enzyme was analyzed by differential scanning calorimetry, which shows a single endothermic transition in the thermogram, with a maximum at 94 °C, while the CD spectra in the visible region indicate the presence of the type 1 copper center up to 80 °C. The reoxidation of the <i>N. gonorrhoeae</i> copper-containing nitrite reductase in the presence of nitrite were analyzed by visible spectroscopy and showed a pH dependence, being higher at pH 5.5–6.0. The high thermostability of this enzyme may be important to maintaining a high activity in the extracellular space and to making it less susceptible to denaturation and proteolysis, contributing to the proliferation of <i>N. gonorrhoeae</i>. |
| first_indexed | 2024-03-11T00:05:36Z |
| format | Article |
| id | doaj.art-9fbfd0e49419491db0c0c243303d259f |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-11T00:05:36Z |
| publishDate | 2023-08-01 |
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| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-9fbfd0e49419491db0c0c243303d259f2023-11-19T00:23:51ZengMDPI AGBiomolecules2218-273X2023-08-01138121510.3390/biom13081215Biochemical Characterization of the Copper Nitrite Reductase from <i>Neisseria gonorrhoeae</i>Daniela S. Barreiro0Ricardo N. S. Oliveira1Sofia R. Pauleta2Microbial Stress Lab, UCIBIO—Applied Molecular Biosciences Unit, Department of Chemistry, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, PortugalMicrobial Stress Lab, UCIBIO—Applied Molecular Biosciences Unit, Department of Chemistry, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, PortugalMicrobial Stress Lab, UCIBIO—Applied Molecular Biosciences Unit, Department of Chemistry, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, PortugalThe copper-containing nitrite reductase from <i>Neisseria gonorrhoeae</i> has been shown to play a critical role in the infection mechanism of this microorganism by producing NO and abolishing epithelial exfoliation. This enzyme is a trimer with a type 1 copper center per subunit and a type 2 copper center in the subunits interface, with the latter being the catalytic site. The two centers were characterized for the first time by EPR and CD spectroscopy, showing that the type 1 copper center has a high rhombicity due to its lower symmetry and more tetragonal structure, while the type 2 copper center has the usual properties, but with a smaller hyperfine coupling constant (A<sub>//</sub> = 10.5 mT). The thermostability of the enzyme was analyzed by differential scanning calorimetry, which shows a single endothermic transition in the thermogram, with a maximum at 94 °C, while the CD spectra in the visible region indicate the presence of the type 1 copper center up to 80 °C. The reoxidation of the <i>N. gonorrhoeae</i> copper-containing nitrite reductase in the presence of nitrite were analyzed by visible spectroscopy and showed a pH dependence, being higher at pH 5.5–6.0. The high thermostability of this enzyme may be important to maintaining a high activity in the extracellular space and to making it less susceptible to denaturation and proteolysis, contributing to the proliferation of <i>N. gonorrhoeae</i>.https://www.mdpi.com/2218-273X/13/8/1215copper nitrite reductaseAniA<i>Neisseria</i>copper enzymenitrite reductionT1 copper center |
| spellingShingle | Daniela S. Barreiro Ricardo N. S. Oliveira Sofia R. Pauleta Biochemical Characterization of the Copper Nitrite Reductase from <i>Neisseria gonorrhoeae</i> Biomolecules copper nitrite reductase AniA <i>Neisseria</i> copper enzyme nitrite reduction T1 copper center |
| title | Biochemical Characterization of the Copper Nitrite Reductase from <i>Neisseria gonorrhoeae</i> |
| title_full | Biochemical Characterization of the Copper Nitrite Reductase from <i>Neisseria gonorrhoeae</i> |
| title_fullStr | Biochemical Characterization of the Copper Nitrite Reductase from <i>Neisseria gonorrhoeae</i> |
| title_full_unstemmed | Biochemical Characterization of the Copper Nitrite Reductase from <i>Neisseria gonorrhoeae</i> |
| title_short | Biochemical Characterization of the Copper Nitrite Reductase from <i>Neisseria gonorrhoeae</i> |
| title_sort | biochemical characterization of the copper nitrite reductase from i neisseria gonorrhoeae i |
| topic | copper nitrite reductase AniA <i>Neisseria</i> copper enzyme nitrite reduction T1 copper center |
| url | https://www.mdpi.com/2218-273X/13/8/1215 |
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