Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Lu, Li-Min, Li, Qing-Zhang, Huang, Jian-Guo, Gao, Xue-Jun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2012
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
_version_ 1783376726150086656
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
work_keys_str_mv AT lulimin proteomicandfunctionalanalysesrevealmapk1regulatesmilkproteinsynthesis
AT liqingzhang proteomicandfunctionalanalysesrevealmapk1regulatesmilkproteinsynthesis
AT huangjianguo proteomicandfunctionalanalysesrevealmapk1regulatesmilkproteinsynthesis
AT gaoxuejun proteomicandfunctionalanalysesrevealmapk1regulatesmilkproteinsynthesis