DICP OpenIR
Subject Area物理化学
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, Shuaibin1,2; Yang, Kaiguang1; Liang, Zhen1; Zhang, Lihua1; Zhang, Yukui1; Zhang LH(张丽华)
KeywordAcid Cleavage Mass Spectrometry Protein Identification Proteomics Sample Preparation
Source PublicationTALANTA
2011-10-30
ISSN待补充
DOI10.1016/j.talanta.2011.08.052
Volume86Issue:待补充Pages:429-435
Indexed BySCI
SubtypeArticle
Department18
Funding Project1810
Contribution Rank1,1
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordLC-ESI-MS/MS ; HYDROLYSIS ; THROUGHPUT ; MIXTURES ; RESIDUES
AbstractUrea free and more efficient sample preparation method for mass spectrometry based protein identification via combining the formic acid-assisted chemical cleavage and trypsin digestion; 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.
Language英语
WOS IDWOS:000298126300057
Citation statistics
Cited Times:6[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/115860
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhang LH(张丽华)
Affiliation1.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
Recommended Citation
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,&张丽华.(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.
Files in This Item:
File Name/Size DocType Version Access License
2011984R1686sk.pdf(687KB) 开放获取--Application Full Text
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[Wu, Shuaibin]'s Articles
[Yang, Kaiguang]'s Articles
[Liang, Zhen]'s Articles
Baidu academic
Similar articles in Baidu academic
[Wu, Shuaibin]'s Articles
[Yang, Kaiguang]'s Articles
[Liang, Zhen]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[Wu, Shuaibin]'s Articles
[Yang, Kaiguang]'s Articles
[Liang, Zhen]'s Articles
Terms of Use
No data!
Social Bookmark/Share
All comments (0)
No comment.
 

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.