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Enrichment of membrane proteins by partitioning in detergent/polymer aqueous two-phase systems.

Everberg, Henrik LU ; Gustavsson, Niklas LU and Tjerneld, Folke LU (2008) In Methods in Molecular Biology 424. p.403-412
Abstract
Methods that combine efficient solubilization with enrichment of proteins and intact protein complexes are of central interest in current membrane proteomics. We have developed methods based on nondenaturing detergent extraction of yeast mitochondrial membrane proteins followed by enrichment of hydrophobic proteins in aqueous two-phase system. Combining the zwitterionic detergent Zwittergent 3-10 and the nonionic detergent Triton X-114 results in a complementary solubilization of proteins, which is similar to that of the anionic detergent sodium dodecyl sulfate (SDS) but with the important advantage of being nondenaturing. Detergent/polymer two-phase system partitioning offers removal of soluble proteins that can be further improved by... (More)
Methods that combine efficient solubilization with enrichment of proteins and intact protein complexes are of central interest in current membrane proteomics. We have developed methods based on nondenaturing detergent extraction of yeast mitochondrial membrane proteins followed by enrichment of hydrophobic proteins in aqueous two-phase system. Combining the zwitterionic detergent Zwittergent 3-10 and the nonionic detergent Triton X-114 results in a complementary solubilization of proteins, which is similar to that of the anionic detergent sodium dodecyl sulfate (SDS) but with the important advantage of being nondenaturing. Detergent/polymer two-phase system partitioning offers removal of soluble proteins that can be further improved by manipulation of the driving forces governing protein distribution between the phases. Integral and peripheral membrane protein subunits from intact membrane protein complexes partition to the detergent phase while soluble proteins are found in the polymer phase. An optimized solubilization protocol is presented in combination with detergent/polymer two-phase partitioning as a mild and efficient method for initial enrichment of membrane proteins and membrane protein complexes in proteomic studies. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Aqueous two-phase systems, detergents, enrichment, membrane proteins, proteomics, solubilization
in
Methods in Molecular Biology
volume
424
pages
403 - 412
publisher
Springer
external identifiers
  • pmid:18369878
  • scopus:44049095537
ISSN
1940-6029
DOI
10.1007/978-1-60327-064-9_31
language
English
LU publication?
yes
id
2b44f166-ffa6-4e25-b02a-1618809ecd7e (old id 1052101)
date added to LUP
2008-05-23 09:16:42
date last changed
2017-01-01 04:32:50
@article{2b44f166-ffa6-4e25-b02a-1618809ecd7e,
  abstract     = {Methods that combine efficient solubilization with enrichment of proteins and intact protein complexes are of central interest in current membrane proteomics. We have developed methods based on nondenaturing detergent extraction of yeast mitochondrial membrane proteins followed by enrichment of hydrophobic proteins in aqueous two-phase system. Combining the zwitterionic detergent Zwittergent 3-10 and the nonionic detergent Triton X-114 results in a complementary solubilization of proteins, which is similar to that of the anionic detergent sodium dodecyl sulfate (SDS) but with the important advantage of being nondenaturing. Detergent/polymer two-phase system partitioning offers removal of soluble proteins that can be further improved by manipulation of the driving forces governing protein distribution between the phases. Integral and peripheral membrane protein subunits from intact membrane protein complexes partition to the detergent phase while soluble proteins are found in the polymer phase. An optimized solubilization protocol is presented in combination with detergent/polymer two-phase partitioning as a mild and efficient method for initial enrichment of membrane proteins and membrane protein complexes in proteomic studies.},
  author       = {Everberg, Henrik and Gustavsson, Niklas and Tjerneld, Folke},
  issn         = {1940-6029},
  keyword      = {Aqueous two-phase systems,detergents,enrichment,membrane proteins,proteomics,solubilization},
  language     = {eng},
  pages        = {403--412},
  publisher    = {Springer},
  series       = {Methods in Molecular Biology},
  title        = {Enrichment of membrane proteins by partitioning in detergent/polymer aqueous two-phase systems.},
  url          = {http://dx.doi.org/10.1007/978-1-60327-064-9_31},
  volume       = {424},
  year         = {2008},
}