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Shotgun analysis of membrane proteomes using a novel combinative strategy of solution-based sample preparation coupled with liquid chromatography-tandem mass spectrometry.

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Shotgun analysis of membrane proteomes using a novel combinative strategy of solution-based sample preparation coupled with liquid chromatography-tandem mass spectrometry.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Lin, Yong Liu, Hui Liu, Zhonghua Wang, Xianchun Liang, Songping
Description	Author Affiliation: Lin Y ( Key Laboratory of Protein Chemistry and Developmental Biology of National Education Committee, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, PR China.)
Abstract	Although much effort has been made in the field of membrane proteomics, the analysis of membrane proteins particularly integral membrane proteins with poor water solubility still presents a great challenge. In this paper, 2% SDS was used to extract membrane proteins and experimental conditions for the application of acetone precipitation method to the cleanup of SDS-solubilized membrane protein sample were optimized. For improving the re-dissolution and trypsinolysis of acetone-precipitated proteins, several commonly used additives, urea, methanol and sodium deoxycholate (SDC), were employed and compared. The results showed that, when the pre-cooled acetone-to-sample ratio was 6:1 (v/v) with one additional washing step, residual SDS in the protein sample could be lowered to below 0.01% and more than 90% of the proteins were precipitated and therefore recovered. 1% SDC-containing buffer could improve the re-dissolution and digestion of the acetone precipitated proteins more efficiently than the others. Using the combinative sample preparation strategy developed, 398 proteins were identified from the rat liver membrane-enriched fraction, including 188 membrane proteins. Compared with other three representative solution-based sample preparation methods commonly used in membrane proteomics, the newly developed combinative strategy increased the number of identified total proteins and membrane proteins on average by 29.2% and 28.5%, respectively. This combinative strategy was demonstrated to be easily operated at low cost and suitable for the analysis of membrane proteins varying in type and sample volume, etc.
File Format	HTM / HTML
ISSN	15700232
Volume Number	901
e-ISSN	1873376X
Journal	Journal of Chromatography B
Language	English
Publisher	Elsevier
Publisher Date	2012-07-15
Publisher Place	Netherlands
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Analytical Chemistry Chromatography, Liquid Methods Membrane Proteins Analysis Peptide Mapping Proteome Tandem Mass Spectrometry Acetone Chemistry Animals Chemical Precipitation Liver Cytology Classification Isolation & Purification Rats Sodium Dodecyl Sulfate Solutions Trypsin Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Cell Biology Medicine Analytical Chemistry Clinical Biochemistry Biochemistry

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Shotgun analysis of membrane proteomes by an improved SDS-assisted sample preparation method coupled with liquid chromatography-tandem mass spectrometry.

Evaluation of the combinative application of SDS and sodium deoxycholate to the LC-MS-based shotgun analysis of membrane proteomes.

Enhanced SDC-assisted digestion coupled with lipid chromatography-tandem mass spectrometry for shotgun analysis of membrane proteome.

Comparison of sodium dodecyl sulfate depletion techniques for proteome analysis by mass spectrometry.

Improvement of a sample preparation method assisted by sodium deoxycholate for mass-spectrometry-based shotgun membrane proteomics.

Comparison of surfactant-assisted shotgun methods using acid-labile surfactants and sodium dodecyl sulfate for membrane proteome analysis.

Universal sample preparation method for proteome analysis.

Sample preparation strategies for targeted proteomics via proteotypic peptides in human blood using liquid chromatography tandem mass spectrometry.

Sample preparation for the analysis of membrane proteomes by mass spectrometry

Shotgun analysis of membrane proteomes using a novel combinative strategy of solution-based sample preparation coupled with liquid chromatography-tandem mass spectrometry.

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Shotgun analysis of membrane proteomes using a novel combinative strategy of solution-based sample preparation coupled with liquid chromatography-tandem mass spectrometry.