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Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System
Nucleoside diphosphate kinase (NDPK, Nm23), a housekeeping enzyme, is known to be a multifunctional protein, acting as a metastasis suppressor, transactivation activity on c-myc, and regulating endocytosis. The cellular mechanisms regulating Nm23 functions are poorly understood. In this study, we id...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2009
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2776532/ https://www.ncbi.nlm.nih.gov/pubmed/19956735 http://dx.doi.org/10.1371/journal.pone.0007949 |
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author | Lee, Eunsun Jeong, Jaeho Kim, Sung Eun Song, Eun Joo Kang, Sang Won Lee, Kong-Joo |
author_facet | Lee, Eunsun Jeong, Jaeho Kim, Sung Eun Song, Eun Joo Kang, Sang Won Lee, Kong-Joo |
author_sort | Lee, Eunsun |
collection | PubMed |
description | Nucleoside diphosphate kinase (NDPK, Nm23), a housekeeping enzyme, is known to be a multifunctional protein, acting as a metastasis suppressor, transactivation activity on c-myc, and regulating endocytosis. The cellular mechanisms regulating Nm23 functions are poorly understood. In this study, we identified the modifications and interacting proteins of Nm23-H1 in response to oxidative stress. We found that Cys109 in Nm23-H1 is oxidized to various oxidation states including intra- and inter-disulfide crosslinks, glutathionylation, and sulfonic acid formation in response to H(2)O(2) treatment both in vivo and in vitro. The cross-linking sites and modifications of oxidized Nm23-H1 were identified by peptide sequencing using UPLC-ESI-q-TOF tandem MS. Glutathionylation and oxidation of Cys109 inhibited the NDPK enzymatic activity of Nm23-H1. We also found that thioredoxin reductase 1 (TrxR1) is an interacting protein of Nm23-H1, and it binds specifically to oxidized Nm23-H1. Oxidized Nm23 is a substrate of NADPH-TrxR1-thioredoxin shuttle system, and the disulfide crosslinking is reversibly reduced and the enzymatic activity is recovered by this system. Oxidation of Cys109 in Nm23-H1 inhibited its metastatic suppressor activity as well as the enzymatic activities. The mutant, Nm23-H1 C109A, retained both the enzymatic and metastasis suppressor activities under oxidative stress. This suggests that key enzymatic and metastasis suppressor functions of Nm23-H1 are regulated by oxido-reduction of its Cys109. |
format | Text |
id | pubmed-2776532 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-27765322009-12-03 Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System Lee, Eunsun Jeong, Jaeho Kim, Sung Eun Song, Eun Joo Kang, Sang Won Lee, Kong-Joo PLoS One Research Article Nucleoside diphosphate kinase (NDPK, Nm23), a housekeeping enzyme, is known to be a multifunctional protein, acting as a metastasis suppressor, transactivation activity on c-myc, and regulating endocytosis. The cellular mechanisms regulating Nm23 functions are poorly understood. In this study, we identified the modifications and interacting proteins of Nm23-H1 in response to oxidative stress. We found that Cys109 in Nm23-H1 is oxidized to various oxidation states including intra- and inter-disulfide crosslinks, glutathionylation, and sulfonic acid formation in response to H(2)O(2) treatment both in vivo and in vitro. The cross-linking sites and modifications of oxidized Nm23-H1 were identified by peptide sequencing using UPLC-ESI-q-TOF tandem MS. Glutathionylation and oxidation of Cys109 inhibited the NDPK enzymatic activity of Nm23-H1. We also found that thioredoxin reductase 1 (TrxR1) is an interacting protein of Nm23-H1, and it binds specifically to oxidized Nm23-H1. Oxidized Nm23 is a substrate of NADPH-TrxR1-thioredoxin shuttle system, and the disulfide crosslinking is reversibly reduced and the enzymatic activity is recovered by this system. Oxidation of Cys109 in Nm23-H1 inhibited its metastatic suppressor activity as well as the enzymatic activities. The mutant, Nm23-H1 C109A, retained both the enzymatic and metastasis suppressor activities under oxidative stress. This suggests that key enzymatic and metastasis suppressor functions of Nm23-H1 are regulated by oxido-reduction of its Cys109. Public Library of Science 2009-11-23 /pmc/articles/PMC2776532/ /pubmed/19956735 http://dx.doi.org/10.1371/journal.pone.0007949 Text en Lee et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Lee, Eunsun Jeong, Jaeho Kim, Sung Eun Song, Eun Joo Kang, Sang Won Lee, Kong-Joo Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System |
title | Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System |
title_full | Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System |
title_fullStr | Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System |
title_full_unstemmed | Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System |
title_short | Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System |
title_sort | multiple functions of nm23-h1 are regulated by oxido-reduction system |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2776532/ https://www.ncbi.nlm.nih.gov/pubmed/19956735 http://dx.doi.org/10.1371/journal.pone.0007949 |
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