An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers

	An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers
	Sui, Zhigang 1; Yuan, Huiming 1; Liang, Zhen 1; Zhao, Qun 1,2; Wu, Qi 1,2; Xia, Simin 1,2; Zhang, Lihua 1; Huo, Yushu 1; Zhang, Yukui 1
刊名	talanta
	2013-03-30
卷号	107 页码:189-194
关键词	Proteome profiling Velvet antlers Sequential protein extraction method Activity assay HUVEC cells
通讯作者	张丽华
产权排序	待补充
合作状况	英
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.
学科主题	物理化学
WOS标题词	science & technology ; physical sciences
类目[WOS]	chemistry, analytical
研究领域[WOS]	chemistry
关键词[WOS]	red deer antler ; expression ; proliferation ; regeneration ; genome ; growth ; tissue ; vegf
收录类别	SCI
资助信息	1，1
原文出处	194
语种	英语
WOS记录号	WOS:000319088700028
公开日期	2014-09-11
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/119107]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
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
推荐引用方式 GB/T 7714	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.
APA	Sui, Zhigang.,Yuan, Huiming.,Liang, Zhen.,Zhao, Qun.,Wu, Qi.,...&Zhang, Yukui.(2013).An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers.talanta,107,189-194.
MLA	Sui, Zhigang,et al."An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers".talanta 107(2013):189-194.