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学科主题: 物理化学
题名: An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers
作者: Sui, Zhigang1;  Yuan, Huiming1;  Liang, Zhen1;  Zhao, Qun1, 2;  Wu, Qi1, 2;  Xia, Simin1, 2;  Zhang, Lihua1;  Huo, Yushu1;  Zhang, Yukui1
通讯作者: 张丽华
关键词: Proteome profiling ;  Velvet antlers ;  Sequential protein extraction method ;  Activity assay ;  HUVEC cells
刊名: TALANTA
发表日期: 2013-03-30
DOI: 10.1016/j.talanta.2013.01.015
卷: 107, 页:189-194
收录类别: SCI
合作性质: 
文章类型: Article
部门归属: 18
项目归属: 1810
产权排名: 待补充
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
资助者: 1,1
研究领域[WOS]: Chemistry
英文摘要: The exceptional growth rate of velvet antler makes it a valuable model for studying the development of tissues, such as blood vessels, cartilage and bone. Meanwhile, investigating the activities of extracted proteins from velvet antlers promisingly leads to the discovery of new active factors which regulate the development of above-mentioned tissue types. In this study, a novel sequential protein extraction method was developed for proteome profiling and bioactivity study of velvet antlers. Herein, four antler growing tips were pooled to create a proportional pooled sample, and three aliquots of which were extracted in parallel using the developed extraction method. For each sample, proteins were extracted sequentially by saline solvent (0.15 M sodium chloride, pH 7.0), mild acid buffer (0.15 M acetate buffer, pH 4.0) and mild alkaline buffer (0.15 M glycine-sodium hydroxide buffer, pH 10.0) with good bio-compatibility to prevent proteins denaturation. Then STD lysis buffer, containing 4% SDS, 0.1 M Tris-HCl and 0.1 M DTT, was used to extract hydrophobic proteins. The tryptic digest of each fraction was analyzed by nanoRPLC-ESI-MS/MS in triplicates, with false discovery rate for peptide identification adjusted to 1% to create filtered protein group list. In total, 1423 protein groups were identified, which expanded up to 3 times of the previous published dataset The relative standard deviation of identified peptide and protein group number for all analyses indicated the good reproducibility of the developed sequential protein extraction method. Additionally, proteins extracted by acid buffer and alkaline buffer showed obvious promoting effect on the proliferation of human umbilical vein endothelial cells. All these results demonstrate that the developed sequential extraction method is efficient for the comprehensive proteome analysis and activity investigation of velvet antlers. (C) 2013 Elsevier B.V. All rights reserved.
关键词[WOS]: RED DEER ANTLER ;  EXPRESSION ;  PROLIFERATION ;  REGENERATION ;  GENOME ;  GROWTH ;  TISSUE ;  VEGF
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000319088700028
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/119107
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100093, Peoples R China

Recommended Citation:
Sui, Zhigang,Yuan, Huiming,Liang, Zhen,et al. An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers[J]. TALANTA,2013,107:189-194.
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