Separation and purification of the bovine milk fat globule membrane protein and its effect on improvement of C2C12 mouse skeletal muscle cell proliferation

2.50
Hdl Handle:
http://hdl.handle.net/10547/622257
Title:
Separation and purification of the bovine milk fat globule membrane protein and its effect on improvement of C2C12 mouse skeletal muscle cell proliferation
Authors:
Li, He; Xu, Weili; Ma, Ying; Zhou, Shaobo ( 0000-0001-5214-2973 )
Abstract:
A novel method to improve the proliferation activity of C2C12 cells by the bovine milk fat globule membrane (MFGM) protein was established in this study. The MFGM protein was extracted and isolated into 4 fractions using an electric cream separator, and purified by a cellulose DEAE-52 column. Fraction 2 accounted for 57.8% of the total MFGM protein, and was used in the following study. The MTT assay showed that it induced cell proliferation activity, increased the cell survival rate and the cell number using flow cytometry and fluorescence microscopy analysis. There were only subtle changes in the morphology as observed using confocal scanning laser microscopy, but the number of mitochondria was significantly increased as observed using transmission electron microscopy analysis. Furthermore, the mRNA expression of MyoD, cyclin D1, p70S6K and mTOR was up-regulated as determined utilizing the quantitative real-time PCR assay, and the activation of Akt and mTOR phosphorylation was up regulated as determined using the Western blot assay. The main protein in fraction 2, assayed by 1-D gel electrophoresis and MALDI TOF-TOF, was identified as milk fat globule-EGF factor 8, the content was 65.6% of the total protein in fraction 2. The results elucidate a new molecular mechanism of the MFGM protein fraction 2: the activation of the Akt signal pathway in promoting cell proliferation.
Affiliation:
Harbin Institute of Technology; University of Bedfordshire
Citation:
Li H, Xu W, Ma Y, Zhou S (2017) 'Separation and purification of the bovine milk fat globule membrane protein and its effect on improvement of C2C12 mouse skeletal muscle cell proliferation', New Journal of Chemistry, 41, pp.6530-6539.
Publisher:
Royal Society of Chemistry
Journal:
New Journal of Chemistry
Issue Date:
7-Jun-2017
URI:
http://hdl.handle.net/10547/622257
DOI:
10.1039/C7NJ00560A
Additional Links:
http://pubs.rsc.org/is/content/articlelanding/2017/nj/c7nj00560a/unauth#!divAbstract
Type:
Article
Language:
en
ISSN:
1144-0546
Sponsors:
National Natural Science Foundation of China grant (NO. 31501481)
Appears in Collections:
Biomedical and biological science

Full metadata record

DC FieldValue Language
dc.contributor.authorLi, Heen
dc.contributor.authorXu, Weilien
dc.contributor.authorMa, Yingen
dc.contributor.authorZhou, Shaoboen
dc.date.accessioned2017-09-28T12:39:13Z-
dc.date.available2017-09-28T12:39:13Z-
dc.date.issued2017-06-07-
dc.identifier.citationLi H, Xu W, Ma Y, Zhou S (2017) 'Separation and purification of the bovine milk fat globule membrane protein and its effect on improvement of C2C12 mouse skeletal muscle cell proliferation', New Journal of Chemistry, 41, pp.6530-6539.en
dc.identifier.issn1144-0546-
dc.identifier.doi10.1039/C7NJ00560A-
dc.identifier.urihttp://hdl.handle.net/10547/622257-
dc.description.abstractA novel method to improve the proliferation activity of C2C12 cells by the bovine milk fat globule membrane (MFGM) protein was established in this study. The MFGM protein was extracted and isolated into 4 fractions using an electric cream separator, and purified by a cellulose DEAE-52 column. Fraction 2 accounted for 57.8% of the total MFGM protein, and was used in the following study. The MTT assay showed that it induced cell proliferation activity, increased the cell survival rate and the cell number using flow cytometry and fluorescence microscopy analysis. There were only subtle changes in the morphology as observed using confocal scanning laser microscopy, but the number of mitochondria was significantly increased as observed using transmission electron microscopy analysis. Furthermore, the mRNA expression of MyoD, cyclin D1, p70S6K and mTOR was up-regulated as determined utilizing the quantitative real-time PCR assay, and the activation of Akt and mTOR phosphorylation was up regulated as determined using the Western blot assay. The main protein in fraction 2, assayed by 1-D gel electrophoresis and MALDI TOF-TOF, was identified as milk fat globule-EGF factor 8, the content was 65.6% of the total protein in fraction 2. The results elucidate a new molecular mechanism of the MFGM protein fraction 2: the activation of the Akt signal pathway in promoting cell proliferation.en
dc.description.sponsorshipNational Natural Science Foundation of China grant (NO. 31501481)en
dc.language.isoenen
dc.publisherRoyal Society of Chemistryen
dc.relation.urlhttp://pubs.rsc.org/is/content/articlelanding/2017/nj/c7nj00560a/unauth#!divAbstracten
dc.rightsGreen - can archive pre-print and post-print or publisher's version/PDF-
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectc2c12en
dc.subjectsacoponiaen
dc.subjectC910 Applied Biological Sciencesen
dc.titleSeparation and purification of the bovine milk fat globule membrane protein and its effect on improvement of C2C12 mouse skeletal muscle cell proliferationen
dc.typeArticleen
dc.contributor.departmentHarbin Institute of Technologyen
dc.contributor.departmentUniversity of Bedfordshireen
dc.identifier.journalNew Journal of Chemistryen
dc.date.updated2017-09-28T12:35:01Z-
dc.description.noteI can provide reprint, if someone needs it. If this is to be eligible for REF, we will need the preprint file (final draft post-refereeing) please! RVO 25-9-17-
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