DICP OpenIR
Subject Area物理化学
Biphasic Microreactor for Efficient Membrane Protein Pretreatment with a Combination of Formic Acid Assisted Solubilization, On-Column pH Adjustment, Reduction, Alkylation, and Tryptic Digestion
Zhao, Qun1,2; Liang, Yu1; Yuan, Huiming1; Sui, Zhigang1; Wu, Qi1,2; Liang, Zhen1; Zhang, Lihua1; Zhang, Yukui1; Zhang LH(张丽华)
Source PublicationANALYTICAL CHEMISTRY
2013-09-17
DOI10.1021/ac402076u
Volume85Issue:18Pages:8507-8512
Indexed BySCI
Cooperation Status
SubtypeArticle
Department18
Funding Project1810
Contribution Rank待补充
WOS HeadingsScience & Technology ; Physical Sciences
Funding Organization1,1 ; 1,1 ; 1,1 ; 1,1
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordTANDEM MASS-SPECTROMETRY ; STRONG-CATION-EXCHANGE ; PHASE LIQUID-CHROMATOGRAPHY ; INTEGRAL MEMBRANE ; PROTEOMIC ANALYSIS ; IDENTIFICATION TECHNOLOGY ; SHOTGUN PROTEOMICS ; TRYPSIN DIGESTION ; BRAIN PROTEOME ; METHANOL
AbstractCombining good dissolving ability of formic acid (FA) for membrane proteins and excellent complementary retention behavior of proteins on strong cation exchange (SCX) and strong anion exchange (SAX) materials, a biphasic microreactor was established to pretreat membrane proteins at microgram and even nanogram levels. With membrane proteins solubilized by FA, all of the proteomics sample processing procedures, including protein preconcentration, pH adjustment, reduction, and alkylation, as well as tryptic digestion, were integrated into an "SCX-SAX" biphasic capillary column. To evaluate the performance of the developed microreactor, a mixture of bovine serum albumin, myoglobin, and cytochrome c was pretreated. Compared with the results obtained by the traditional in-solution process, the peptide recovery (93% vs 83%) and analysis throughput (3.5 vs 14 h) were obviously improved. The microreactor was further applied for the pretreatment of 14 mu g of membrane proteins extracted from rat cerebellums, and 416 integral membrane proteins (IMPs) (43% of total protein groups) and 103 transmembrane peptides were identified by two-dimensional nanoliquid chromatography-electrospray ionization tandem mass spectrometry (2D nano-LC-ESI-MS/MS) in triplicate analysis. With the starting sample preparation amount decreased to as low as 50 ng, 64 IMPs and 17 transmembrane peptides were identified confidently, while those obtained by the traditional in-solution method were 10 and 1, respectively. All these results demonstrated that such an "SCX-SAX" based biphasic microreactor could offer a promising tool for the pretreatment of trace membrane proteins with high efficiency and throughput.
Language英语
Funding Organization1,1 ; 1,1 ; 1,1 ; 1,1
URL查看原文
WOS IDWOS:000330017400007
Citation statistics
Cited Times:13[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/119507
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, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100039, Peoples R China
Recommended Citation
GB/T 7714
Zhao, Qun,Liang, Yu,Yuan, Huiming,et al. Biphasic Microreactor for Efficient Membrane Protein Pretreatment with a Combination of Formic Acid Assisted Solubilization, On-Column pH Adjustment, Reduction, Alkylation, and Tryptic Digestion[J]. ANALYTICAL CHEMISTRY,2013,85(18):8507-8512.
APA Zhao, Qun.,Liang, Yu.,Yuan, Huiming.,Sui, Zhigang.,Wu, Qi.,...&张丽华.(2013).Biphasic Microreactor for Efficient Membrane Protein Pretreatment with a Combination of Formic Acid Assisted Solubilization, On-Column pH Adjustment, Reduction, Alkylation, and Tryptic Digestion.ANALYTICAL CHEMISTRY,85(18),8507-8512.
MLA Zhao, Qun,et al."Biphasic Microreactor for Efficient Membrane Protein Pretreatment with a Combination of Formic Acid Assisted Solubilization, On-Column pH Adjustment, Reduction, Alkylation, and Tryptic Digestion".ANALYTICAL CHEMISTRY 85.18(2013):8507-8512.
Files in This Item:
File Name/Size DocType Version Access License
2013R6XGR629CB.PDF(1132KB) 开放获取CC BY-NC-SAApplication Full Text
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[Zhao, Qun]'s Articles
[Liang, Yu]'s Articles
[Yuan, Huiming]'s Articles
Baidu academic
Similar articles in Baidu academic
[Zhao, Qun]'s Articles
[Liang, Yu]'s Articles
[Yuan, Huiming]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[Zhao, Qun]'s Articles
[Liang, Yu]'s Articles
[Yuan, Huiming]'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.