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Proteomic and Functional Analyses Reveal MAPK1 Regulates Milk Protein Synthesis
L-Lysine (L-Lys) is an essential amino acid that plays fundamental roles in protein synthesis. Many nuclear phosphorylated proteins such as Stat5 and mTOR regulate milk protein synthesis. However, the details of milk protein synthesis control at the transcript and translational levels are not well k...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6270553/ https://www.ncbi.nlm.nih.gov/pubmed/23271465 http://dx.doi.org/10.3390/molecules18010263 |
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author | Lu, Li-Min Li, Qing-Zhang Huang, Jian-Guo Gao, Xue-Jun |
author_facet | Lu, Li-Min Li, Qing-Zhang Huang, Jian-Guo Gao, Xue-Jun |
author_sort | Lu, Li-Min |
collection | PubMed |
description | L-Lysine (L-Lys) is an essential amino acid that plays fundamental roles in protein synthesis. Many nuclear phosphorylated proteins such as Stat5 and mTOR regulate milk protein synthesis. However, the details of milk protein synthesis control at the transcript and translational levels are not well known. In this current study, a two-dimensional gel electrophoresis (2-DE)/MS-based proteomic technology was used to identify phosphoproteins responsible for milk protein synthesis in dairy cow mammary epithelial cells (DCMECs). The effect of L-Lys on DCMECs was analyzed by CASY technology and reversed phase high performance liquid chromatography (RP-HPLC). The results showed that cell proliferation ability and β-casein expression were enhanced in DCMECs treated with L-Lys. By phosphoproteomics analysis, six proteins, including MAPK1, were identified up-expressed in DCMECs treated with 1.2 mM L-Lys for 24 h, and were verified by quantitative real-time PCR (qRT-PCR) and western blot. Overexpression and siRNA inhibition of MAPK1 experiments showed that MAPK1 upregulated milk protein synthesis through Stat5 and mTOR pathway. These findings that MAPK1 involves in regulation of milk synthesis shed new insights for understanding the mechanisms of milk protein synthesis. |
format | Online Article Text |
id | pubmed-6270553 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-62705532018-12-14 Proteomic and Functional Analyses Reveal MAPK1 Regulates Milk Protein Synthesis Lu, Li-Min Li, Qing-Zhang Huang, Jian-Guo Gao, Xue-Jun Molecules Article L-Lysine (L-Lys) is an essential amino acid that plays fundamental roles in protein synthesis. Many nuclear phosphorylated proteins such as Stat5 and mTOR regulate milk protein synthesis. However, the details of milk protein synthesis control at the transcript and translational levels are not well known. In this current study, a two-dimensional gel electrophoresis (2-DE)/MS-based proteomic technology was used to identify phosphoproteins responsible for milk protein synthesis in dairy cow mammary epithelial cells (DCMECs). The effect of L-Lys on DCMECs was analyzed by CASY technology and reversed phase high performance liquid chromatography (RP-HPLC). The results showed that cell proliferation ability and β-casein expression were enhanced in DCMECs treated with L-Lys. By phosphoproteomics analysis, six proteins, including MAPK1, were identified up-expressed in DCMECs treated with 1.2 mM L-Lys for 24 h, and were verified by quantitative real-time PCR (qRT-PCR) and western blot. Overexpression and siRNA inhibition of MAPK1 experiments showed that MAPK1 upregulated milk protein synthesis through Stat5 and mTOR pathway. These findings that MAPK1 involves in regulation of milk synthesis shed new insights for understanding the mechanisms of milk protein synthesis. MDPI 2012-12-27 /pmc/articles/PMC6270553/ /pubmed/23271465 http://dx.doi.org/10.3390/molecules18010263 Text en © 2013 by the authors; licensee MDPI, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0/ This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/). |
spellingShingle | Article Lu, Li-Min Li, Qing-Zhang Huang, Jian-Guo Gao, Xue-Jun Proteomic and Functional Analyses Reveal MAPK1 Regulates Milk Protein Synthesis |
title | Proteomic and Functional Analyses Reveal MAPK1 Regulates Milk Protein Synthesis |
title_full | Proteomic and Functional Analyses Reveal MAPK1 Regulates Milk Protein Synthesis |
title_fullStr | Proteomic and Functional Analyses Reveal MAPK1 Regulates Milk Protein Synthesis |
title_full_unstemmed | Proteomic and Functional Analyses Reveal MAPK1 Regulates Milk Protein Synthesis |
title_short | Proteomic and Functional Analyses Reveal MAPK1 Regulates Milk Protein Synthesis |
title_sort | proteomic and functional analyses reveal mapk1 regulates milk protein synthesis |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6270553/ https://www.ncbi.nlm.nih.gov/pubmed/23271465 http://dx.doi.org/10.3390/molecules18010263 |
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