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Efficient and non-denaturing membrane solubilization combined with enrichment of membrane protein complexes by detergent/polymer aqueous two-phase partitioning for proteome analysis

Everberg, Henrik LU ; Leiding, Thom LU ; Schioth, Anna ; Tjerneld, Folke LU and Gustavsson, Niklas LU (2006) In Journal of Chromatography A 1122(1-2). p.35-46
Abstract
It is of central interest in membrane proteomics to establish methods that combine efficient solubilization with enrichment of proteins and intact protein complexes. We have investigated the quantitative and qualitative solubilization efficiency of five commercially available detergents using mitochondria from the yeast Saccharomyces cerevisiae as model system. Combining the zwitterionic detergent Zwittergent 3-10 and the non-ionic detergent Triton X-114 resulted in a complementary solubilization of proteins, which was similar to that of the anionic detergent sodium dodecyl sulfate (SDS). The subsequent removal of soluble proteins by detergent/polymer two-phase system partitioning was further enhanced by addition of SDS and increasing pH.... (More)
It is of central interest in membrane proteomics to establish methods that combine efficient solubilization with enrichment of proteins and intact protein complexes. We have investigated the quantitative and qualitative solubilization efficiency of five commercially available detergents using mitochondria from the yeast Saccharomyces cerevisiae as model system. Combining the zwitterionic detergent Zwittergent 3-10 and the non-ionic detergent Triton X-114 resulted in a complementary solubilization of proteins, which was similar to that of the anionic detergent sodium dodecyl sulfate (SDS). The subsequent removal of soluble proteins by detergent/polymer two-phase system partitioning was further enhanced by addition of SDS and increasing pH. A large number of both integral and peripheral membrane protein subunits from mitochondrial membrane protein complexes were identified in the detergent phase. We suggest that the optimized solubilization protocol in combination with detergent/polymer two-phase partitioning is a mild and efficient method for initial enrichment of membrane proteins and membrane protein complexes in proteomic studies. (c) 2006 Elsevier B.V. All rights reserved. (Less)
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author
; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
aqueous, membrane proteins, enrichment, detergents, solubilization, proteomics, two-phase systems
in
Journal of Chromatography A
volume
1122
issue
1-2
pages
35 - 46
publisher
Elsevier
external identifiers
  • pmid:16682048
  • wos:000239155300006
  • scopus:33745889633
ISSN
0021-9673
DOI
10.1016/j.chroma.2006.04.020
language
English
LU publication?
yes
id
f53c010a-e1d0-40e1-b96e-f0b16f242759 (old id 401321)
date added to LUP
2016-04-01 15:24:53
date last changed
2021-02-17 01:34:02
@article{f53c010a-e1d0-40e1-b96e-f0b16f242759,
  abstract     = {It is of central interest in membrane proteomics to establish methods that combine efficient solubilization with enrichment of proteins and intact protein complexes. We have investigated the quantitative and qualitative solubilization efficiency of five commercially available detergents using mitochondria from the yeast Saccharomyces cerevisiae as model system. Combining the zwitterionic detergent Zwittergent 3-10 and the non-ionic detergent Triton X-114 resulted in a complementary solubilization of proteins, which was similar to that of the anionic detergent sodium dodecyl sulfate (SDS). The subsequent removal of soluble proteins by detergent/polymer two-phase system partitioning was further enhanced by addition of SDS and increasing pH. A large number of both integral and peripheral membrane protein subunits from mitochondrial membrane protein complexes were identified in the detergent phase. We suggest that the optimized solubilization protocol in combination with detergent/polymer two-phase partitioning is a mild and efficient method for initial enrichment of membrane proteins and membrane protein complexes in proteomic studies. (c) 2006 Elsevier B.V. All rights reserved.},
  author       = {Everberg, Henrik and Leiding, Thom and Schioth, Anna and Tjerneld, Folke and Gustavsson, Niklas},
  issn         = {0021-9673},
  language     = {eng},
  number       = {1-2},
  pages        = {35--46},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Efficient and non-denaturing membrane solubilization combined with enrichment of membrane protein complexes by detergent/polymer aqueous two-phase partitioning for proteome analysis},
  url          = {http://dx.doi.org/10.1016/j.chroma.2006.04.020},
  doi          = {10.1016/j.chroma.2006.04.020},
  volume       = {1122},
  year         = {2006},
}