A sample preparation method for micro-scale membrane
proteome analysis

• Authors: Jianjun Li , Hai Yu , Zhigui Duan , Xianchun Wang
• Languages of publication: EN

Abstracts

EN

Detergents are widely used to improve the solubilization and extraction of
hydrophobic membrane proteins in proteomics. Since most detergents
are not compatible with subsequent steps of analysis, the removal of
detergents from samples, especially those in micro-scale amounts,
is a worthy topic of investigation. In this paper, we present a novel
polyvinylidene difluoride (PVDF) membrane-mediated sample preparation
method for micro-scale membrane proteome analysis, using a rat liver
cell membrane-enriched fraction as model material. The proteins in the
fraction were extracted in a 2 % sodium dodecyl sulfate (SDS) solution and
the protein solution was applied on a piece of PVDF membrane followed
by drying and repeated washing in order to remove SDS and other salts.
Quantitative determination indicated that about 84% of the SDS in the
sample was removed and protein loss was less than 10%. Four methods
were used and compared for digesting the proteins adsorbed on PVDF
membrane. Dimethyl formamide (DMF)-assisted digestion was the most
effective with regard to the identification of membrane proteins, particularly
the highly hydrophobic multi-transmembrane proteins. These results
demonstrate that PVDF membrane-aided sample cleanup combined with
DMF-assisted digestion has potential utility in the micro-scale membrane
proteome analysis.

Keywords

EN

Dimethyl formamide ; Membrane proteome ; Proteomics ; Polyvinylidene difluoride membrane ; SDS

• Publisher: De Gruyter Open
• Journal: Sample Preparation
• Year: 2013
• Volume: 1
• Pages: 28-37

Dates

received

09 - 2 - 2013

online

22 - 11 - 2013

accepted

24 - 08 - 2013

Contributors

author

Jianjun Li

• Key Laboratory of Protein Chemistry
  and Developmental Biology of National
  Education Committee, College of Life
  Sciences, Hunan Normal University,
  Changsha, 410081, P. R. China

author

Hai Yu

• Key Laboratory of Protein Chemistry
  and Developmental Biology of National
  Education Committee, College of Life
  Sciences, Hunan Normal University,
  Changsha, 410081, P. R. China

author

Zhigui Duan

• Key Laboratory of Protein Chemistry
  and Developmental Biology of National
  Education Committee, College of Life
  Sciences, Hunan Normal University,
  Changsha, 410081, P. R. China

author

Xianchun Wang

• Key Laboratory of Protein Chemistry
  and Developmental Biology of National
  Education Committee, College of Life
  Sciences, Hunan Normal University,
  Changsha, 410081, P. R. China

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Identifiers

DOI

10.2478/sampre-2013-0004