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Quantification of Membrane Proteins Using Nonspecific Protease Digestions

Bendz, Maria ; Carlsson Möller, Mirja LU ; Arrigoni, Giorgio LU ; Wåhlander, Åsa LU ; Stella, Roberto ; Cappadona, Salvatore LU ; Levander, Fredrik LU ; Hederstedt, Lars LU and James, Peter LU orcid (2009) In Journal of Proteome Research 8(12). p.5666-5673
Abstract
We present a mass spectrometry-based method for the identification and quantification of membrane proteins using the low-specificity protease Proteinase K, at very high pH, to digest proteins isolated by a modified SDS-PAGE protocol. The resulting peptides are modified with a fragmentation-directing isotope labeled tag. We apply the method to quantify differences in membrane protein expression of Bacillus subtilis grown in the presence or absence of glucose.
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author
; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
membrane proteins, N-terminal labeling, quantification, relative, proteinase K, Bacillus subtilis
in
Journal of Proteome Research
volume
8
issue
12
pages
5666 - 5673
publisher
The American Chemical Society (ACS)
external identifiers
  • wos:000272339100026
  • scopus:71549132222
ISSN
1535-3893
DOI
10.1021/pr900741t
language
English
LU publication?
yes
id
b5889499-14d3-4b76-b0a7-097924af96a8 (old id 1533438)
date added to LUP
2016-04-01 12:38:08
date last changed
2023-11-17 13:00:03
@article{b5889499-14d3-4b76-b0a7-097924af96a8,
  abstract     = {{We present a mass spectrometry-based method for the identification and quantification of membrane proteins using the low-specificity protease Proteinase K, at very high pH, to digest proteins isolated by a modified SDS-PAGE protocol. The resulting peptides are modified with a fragmentation-directing isotope labeled tag. We apply the method to quantify differences in membrane protein expression of Bacillus subtilis grown in the presence or absence of glucose.}},
  author       = {{Bendz, Maria and Carlsson Möller, Mirja and Arrigoni, Giorgio and Wåhlander, Åsa and Stella, Roberto and Cappadona, Salvatore and Levander, Fredrik and Hederstedt, Lars and James, Peter}},
  issn         = {{1535-3893}},
  keywords     = {{membrane proteins; N-terminal labeling; quantification; relative; proteinase K; Bacillus subtilis}},
  language     = {{eng}},
  number       = {{12}},
  pages        = {{5666--5673}},
  publisher    = {{The American Chemical Society (ACS)}},
  series       = {{Journal of Proteome Research}},
  title        = {{Quantification of Membrane Proteins Using Nonspecific Protease Digestions}},
  url          = {{http://dx.doi.org/10.1021/pr900741t}},
  doi          = {{10.1021/pr900741t}},
  volume       = {{8}},
  year         = {{2009}},
}