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Insights into the Role of Substrates on the Interaction between Cytochrome b(5) and Cytochrome P450 2B4 by NMR
Mammalian cytochrome b(5) (cyt b(5)) is a membrane-bound protein capable of donating an electron to cytochrome P450 (P450) in the P450 catalytic cycle. The interaction between cyt b(5) and P450 has been reported to be affected by the substrates of P450; however, the mechanism of substrate modulation...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4330534/ https://www.ncbi.nlm.nih.gov/pubmed/25687717 http://dx.doi.org/10.1038/srep08392 |
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author | Zhang, Meng Le Clair, Stéphanie V. Huang, Rui Ahuja, Shivani Im, Sang-Choul Waskell, Lucy Ramamoorthy, Ayyalusamy |
author_facet | Zhang, Meng Le Clair, Stéphanie V. Huang, Rui Ahuja, Shivani Im, Sang-Choul Waskell, Lucy Ramamoorthy, Ayyalusamy |
author_sort | Zhang, Meng |
collection | PubMed |
description | Mammalian cytochrome b(5) (cyt b(5)) is a membrane-bound protein capable of donating an electron to cytochrome P450 (P450) in the P450 catalytic cycle. The interaction between cyt b(5) and P450 has been reported to be affected by the substrates of P450; however, the mechanism of substrate modulation on the cyt b(5)-P450 complex formation is still unknown. In this study, the complexes between full-length rabbit cyt b(5) and full-length substrate-free/substrate-bound cytochrome P450 2B4 (CYP2B4) are investigated using NMR techniques. Our findings reveal that the population of complexes is ionic strength dependent, implying the importance of electrostatic interactions in the complex formation process. The observation that the cyt b(5)-substrate-bound CYP2B4 complex shows a weaker dependence on ionic strength than the cyt b(5)-substrate-free CYP2B4 complex suggests the presence of a larger fraction of steoreospecific complexes when CYP2B4 is substrate-bound. These results suggest that a CYP2B4 substrate likely promotes specific interactions between cyt b(5) and CYP2B4. Residues D65, V66, T70, D71 and A72 are found to be involved in specific interactions between the two proteins due to their weak response to ionic strength change. These findings provide insights into the mechanism underlying substrate modulation on the cyt b(5)-P450 complexation process. |
format | Online Article Text |
id | pubmed-4330534 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-43305342015-02-23 Insights into the Role of Substrates on the Interaction between Cytochrome b(5) and Cytochrome P450 2B4 by NMR Zhang, Meng Le Clair, Stéphanie V. Huang, Rui Ahuja, Shivani Im, Sang-Choul Waskell, Lucy Ramamoorthy, Ayyalusamy Sci Rep Article Mammalian cytochrome b(5) (cyt b(5)) is a membrane-bound protein capable of donating an electron to cytochrome P450 (P450) in the P450 catalytic cycle. The interaction between cyt b(5) and P450 has been reported to be affected by the substrates of P450; however, the mechanism of substrate modulation on the cyt b(5)-P450 complex formation is still unknown. In this study, the complexes between full-length rabbit cyt b(5) and full-length substrate-free/substrate-bound cytochrome P450 2B4 (CYP2B4) are investigated using NMR techniques. Our findings reveal that the population of complexes is ionic strength dependent, implying the importance of electrostatic interactions in the complex formation process. The observation that the cyt b(5)-substrate-bound CYP2B4 complex shows a weaker dependence on ionic strength than the cyt b(5)-substrate-free CYP2B4 complex suggests the presence of a larger fraction of steoreospecific complexes when CYP2B4 is substrate-bound. These results suggest that a CYP2B4 substrate likely promotes specific interactions between cyt b(5) and CYP2B4. Residues D65, V66, T70, D71 and A72 are found to be involved in specific interactions between the two proteins due to their weak response to ionic strength change. These findings provide insights into the mechanism underlying substrate modulation on the cyt b(5)-P450 complexation process. Nature Publishing Group 2015-02-17 /pmc/articles/PMC4330534/ /pubmed/25687717 http://dx.doi.org/10.1038/srep08392 Text en Copyright © 2015, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Zhang, Meng Le Clair, Stéphanie V. Huang, Rui Ahuja, Shivani Im, Sang-Choul Waskell, Lucy Ramamoorthy, Ayyalusamy Insights into the Role of Substrates on the Interaction between Cytochrome b(5) and Cytochrome P450 2B4 by NMR |
title | Insights into the Role of Substrates on the Interaction between Cytochrome b(5) and Cytochrome P450 2B4 by NMR |
title_full | Insights into the Role of Substrates on the Interaction between Cytochrome b(5) and Cytochrome P450 2B4 by NMR |
title_fullStr | Insights into the Role of Substrates on the Interaction between Cytochrome b(5) and Cytochrome P450 2B4 by NMR |
title_full_unstemmed | Insights into the Role of Substrates on the Interaction between Cytochrome b(5) and Cytochrome P450 2B4 by NMR |
title_short | Insights into the Role of Substrates on the Interaction between Cytochrome b(5) and Cytochrome P450 2B4 by NMR |
title_sort | insights into the role of substrates on the interaction between cytochrome b(5) and cytochrome p450 2b4 by nmr |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4330534/ https://www.ncbi.nlm.nih.gov/pubmed/25687717 http://dx.doi.org/10.1038/srep08392 |
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