Urea free and more efficient sample preparation method for mass spectrometry based protein identification via combining the formic acid-assisted chemical cleavage and trypsin digestion

	Urea free and more efficient sample preparation method for mass spectrometry based protein identification via combining the formic acid-assisted chemical cleavage and trypsin digestion
	Wu, Shuaibin 1,2; Yang, Kaiguang 1; Liang, Zhen 1; Zhang, Lihua 1; Zhang, Yukui 1
刊名	talanta
	2011-10-30
卷号	86 期号:待补充页码:429-435
关键词	Acid cleavage Mass spectrometry Protein identification Proteomics Sample preparation
ISSN号	待补充
通讯作者	张丽华
产权排序	1,1
中文摘要	urea free and more efficient sample preparation method for mass spectrometry based protein identification via combining the formic acid-assisted chemical cleavage and trypsin digestion
英文摘要	a formic acid (fa)-assisted sample preparation method was presented for protein identification via mass spectrometry (ms). detailedly, an aqueous solution containing 2% fa and dithiothreitol was selected to perform protein denaturation, aspartic acid (d) sites cleavage and disulfide linkages reduction simultaneously at 108 degrees c for 2h. subsequently. fa wiped off via vacuum concentration. finally, iodoacetamide (iaa) alkylation and trypsin digestion could be performed ordinally. a series of model proteins (bsa. beta-lactoglobulin and apo-transferrin) were treated respectively using such method, followed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (maldi-tof ms) analysis. the identified peptide number was increased by similar to 80% in comparison with the conventional urea-assisted sample preparation method. moreover, bsa identification was achieved efficiently down to femtomole (25 +/- 0 sequence coverage and 16 +/- 1 peptides) via such method. in contrast, there were not peptides identified confidently via the urea-assisted method before desalination via the c18 zip tip. the absence of urea in this sample preparation method was an advantage for the more favorable digestion and maldi-tof ms analysis. the performances of two methods for the real sample (rat liver proteome) were also compared, followed by a nanoflow reversed-phase liquid chromatography with electrospray ionization tandem mass spectrometry system analysis. as a result, 1335 +/- 43 peptides were identified confidently (false discovery rate <1%) via fa-assisted method, corresponding to 295 +/- 12 proteins (of top match = 1 and requiring 2 unique peptides at least). in contrast, there were only 1107 +/- 16 peptides (corresponding to 231 +/- 10 proteins) obtained from the conventional urea-assisted method. it was serving as a more efficient protein sample preparation method for researching specific proteomes better, and providing assistance to develop other proteomics analysis methods, such as, peptide quantitative analysis. (c) 2011 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]	lc-esi-ms/ms ; hydrolysis ; throughput ; mixtures ; residues
收录类别	SCI
语种	英语
WOS记录号	WOS:000298126300057
公开日期	2012-07-09
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/115860]
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.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Wu, Shuaibin,Yang, Kaiguang,Liang, Zhen,et al. Urea free and more efficient sample preparation method for mass spectrometry based protein identification via combining the formic acid-assisted chemical cleavage and trypsin digestion[J]. talanta,2011,86(待补充):429-435.
APA	Wu, Shuaibin,Yang, Kaiguang,Liang, Zhen,Zhang, Lihua,&Zhang, Yukui.(2011).Urea free and more efficient sample preparation method for mass spectrometry based protein identification via combining the formic acid-assisted chemical cleavage and trypsin digestion.talanta,86(待补充),429-435.
MLA	Wu, Shuaibin,et al."Urea free and more efficient sample preparation method for mass spectrometry based protein identification via combining the formic acid-assisted chemical cleavage and trypsin digestion".talanta 86.待补充(2011):429-435.