Changes in Hemoglobin Properties in Complex with Glutathione and after Glutathionylation
Hemoglobin is the main protein of red blood cells that provides oxygen transport to all cells of the human body. The ability of hemoglobin to bind the main low-molecular-weight thiol of the cell glutathione, both covalently and noncovalently, is not only an important part of the antioxidant protecti...
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2023-08-01
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| Series: | International Journal of Molecular Sciences |
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| author | Iuliia D. Kuleshova Pavel I. Zaripov Yuri M. Poluektov Anastasia A. Anashkina Dmitry N. Kaluzhny Evgeniia Yu. Parshina Georgy V. Maksimov Vladimir A. Mitkevich Alexander A. Makarov Irina Yu. Petrushanko |
| author_facet | Iuliia D. Kuleshova Pavel I. Zaripov Yuri M. Poluektov Anastasia A. Anashkina Dmitry N. Kaluzhny Evgeniia Yu. Parshina Georgy V. Maksimov Vladimir A. Mitkevich Alexander A. Makarov Irina Yu. Petrushanko |
| author_sort | Iuliia D. Kuleshova |
| collection | DOAJ |
| description | Hemoglobin is the main protein of red blood cells that provides oxygen transport to all cells of the human body. The ability of hemoglobin to bind the main low-molecular-weight thiol of the cell glutathione, both covalently and noncovalently, is not only an important part of the antioxidant protection of red blood cells, but also affects its affinity for oxygen in both cases. In this study, the properties of oxyhemoglobin in complex with reduced glutathione (GSH) and properties of glutathionylated hemoglobin bound to glutathione via an SS bond were characterized. For this purpose, the methods of circular dichroism, Raman spectroscopy, infrared spectroscopy, tryptophan fluorescence, differential scanning fluorimetry, and molecular modeling were used. It was found that the glutathionylation of oxyhemoglobin caused changes in the secondary structure of the protein, reducing the alpha helicity, but did not affect the heme environment, tryptophan fluorescence, and the thermostability of the protein. In the noncovalent complex of oxyhemoglobin with reduced glutathione, the secondary structure of hemoglobin remained almost unchanged; however, changes in the heme environment and the microenvironment of tryptophans, as well as a decrease in the protein’s thermal stability, were observed. Thus, the formation of a noncovalent complex of hemoglobin with glutathione makes a more significant effect on the tertiary and quaternary structure of hemoglobin than glutathionylation, which mainly affects the secondary structure of the protein. The obtained data are important for understanding the functioning of glutathionylated hemoglobin, which is a marker of oxidative stress, and hemoglobin in complex with GSH, which appears to deposit GSH and release it during deoxygenation to increase the antioxidant protection of cells. |
| first_indexed | 2024-03-10T23:20:21Z |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T23:20:21Z |
| publishDate | 2023-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-473b8474eea04ca58e48eed1e135c5122023-11-19T08:18:59ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-08-0124171355710.3390/ijms241713557Changes in Hemoglobin Properties in Complex with Glutathione and after GlutathionylationIuliia D. Kuleshova0Pavel I. Zaripov1Yuri M. Poluektov2Anastasia A. Anashkina3Dmitry N. Kaluzhny4Evgeniia Yu. Parshina5Georgy V. Maksimov6Vladimir A. Mitkevich7Alexander A. Makarov8Irina Yu. Petrushanko9Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, RussiaEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, RussiaEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, RussiaEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, RussiaEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, RussiaFaculty of Biology, Lomonosov Moscow State University, Moscow 119234, RussiaFaculty of Biology, Lomonosov Moscow State University, Moscow 119234, RussiaEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, RussiaEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, RussiaEngelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow 119991, RussiaHemoglobin is the main protein of red blood cells that provides oxygen transport to all cells of the human body. The ability of hemoglobin to bind the main low-molecular-weight thiol of the cell glutathione, both covalently and noncovalently, is not only an important part of the antioxidant protection of red blood cells, but also affects its affinity for oxygen in both cases. In this study, the properties of oxyhemoglobin in complex with reduced glutathione (GSH) and properties of glutathionylated hemoglobin bound to glutathione via an SS bond were characterized. For this purpose, the methods of circular dichroism, Raman spectroscopy, infrared spectroscopy, tryptophan fluorescence, differential scanning fluorimetry, and molecular modeling were used. It was found that the glutathionylation of oxyhemoglobin caused changes in the secondary structure of the protein, reducing the alpha helicity, but did not affect the heme environment, tryptophan fluorescence, and the thermostability of the protein. In the noncovalent complex of oxyhemoglobin with reduced glutathione, the secondary structure of hemoglobin remained almost unchanged; however, changes in the heme environment and the microenvironment of tryptophans, as well as a decrease in the protein’s thermal stability, were observed. Thus, the formation of a noncovalent complex of hemoglobin with glutathione makes a more significant effect on the tertiary and quaternary structure of hemoglobin than glutathionylation, which mainly affects the secondary structure of the protein. The obtained data are important for understanding the functioning of glutathionylated hemoglobin, which is a marker of oxidative stress, and hemoglobin in complex with GSH, which appears to deposit GSH and release it during deoxygenation to increase the antioxidant protection of cells.https://www.mdpi.com/1422-0067/24/17/13557hemoglobinglutathioneglutathionylationcircular dichroismRaman scatteringinfrared spectroscopy |
| spellingShingle | Iuliia D. Kuleshova Pavel I. Zaripov Yuri M. Poluektov Anastasia A. Anashkina Dmitry N. Kaluzhny Evgeniia Yu. Parshina Georgy V. Maksimov Vladimir A. Mitkevich Alexander A. Makarov Irina Yu. Petrushanko Changes in Hemoglobin Properties in Complex with Glutathione and after Glutathionylation International Journal of Molecular Sciences hemoglobin glutathione glutathionylation circular dichroism Raman scattering infrared spectroscopy |
| title | Changes in Hemoglobin Properties in Complex with Glutathione and after Glutathionylation |
| title_full | Changes in Hemoglobin Properties in Complex with Glutathione and after Glutathionylation |
| title_fullStr | Changes in Hemoglobin Properties in Complex with Glutathione and after Glutathionylation |
| title_full_unstemmed | Changes in Hemoglobin Properties in Complex with Glutathione and after Glutathionylation |
| title_short | Changes in Hemoglobin Properties in Complex with Glutathione and after Glutathionylation |
| title_sort | changes in hemoglobin properties in complex with glutathione and after glutathionylation |
| topic | hemoglobin glutathione glutathionylation circular dichroism Raman scattering infrared spectroscopy |
| url | https://www.mdpi.com/1422-0067/24/17/13557 |
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