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Quantitative Assessment of Urea In-Solution Lys-C/Trypsin Digestions Reveals Superior Performance at Room Temperature over Traditional Proteolysis at 37 °C.

Betancourt, Lazaro LU ; Sanchez, Aniel LU ; Pla Parada, Indira LU ; Kuras, Magdalena LU ; Zhou, Qimin LU ; Andersson, Roland LU and Marko-Varga, György LU (2018) In Journal of Proteome Research 17(7). p.2556-2561
Abstract
Urea-containing buffer solutions are generally used in proteomic studies to aid protein denaturation and solubilization during cell and tissue lysis. It is well-known, however, that urea can lead to carbamylation of peptides and proteins and, subsequently, incomplete digestion of proteins. By the use of cells and tissues that had been lysed with urea, different solution digestion strategies were quantitatively assessed. In comparison with traditional proteolysis at 37 °C, urea in-solution digestion performed at room temperature improved peptide and protein identification and quantitation and had a minimum impact on miscleavage rates. Furthermore, the signal intensities and the number of carbamylated and pyroglutamic acid-modified peptides... (More)
Urea-containing buffer solutions are generally used in proteomic studies to aid protein denaturation and solubilization during cell and tissue lysis. It is well-known, however, that urea can lead to carbamylation of peptides and proteins and, subsequently, incomplete digestion of proteins. By the use of cells and tissues that had been lysed with urea, different solution digestion strategies were quantitatively assessed. In comparison with traditional proteolysis at 37 °C, urea in-solution digestion performed at room temperature improved peptide and protein identification and quantitation and had a minimum impact on miscleavage rates. Furthermore, the signal intensities and the number of carbamylated and pyroglutamic acid-modified peptides decreased. Overall, this led to a reduction in the negative effects often observed for such modifications. Data are available via ProteomeXchange with identifier PXD009426. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Urea, proteomics
in
Journal of Proteome Research
volume
17
issue
7
pages
6 pages
publisher
The American Chemical Society
external identifiers
  • scopus:85047969180
ISSN
1535-3893
DOI
10.1021/acs.jproteome.8b00228
language
English
LU publication?
yes
id
1fdd9bb8-afc7-47c9-8519-21ee5f6fb7f6
date added to LUP
2018-12-13 15:07:46
date last changed
2019-03-19 04:03:30
@article{1fdd9bb8-afc7-47c9-8519-21ee5f6fb7f6,
  abstract     = {Urea-containing buffer solutions are generally used in proteomic studies to aid protein denaturation and solubilization during cell and tissue lysis. It is well-known, however, that urea can lead to carbamylation of peptides and proteins and, subsequently, incomplete digestion of proteins. By the use of cells and tissues that had been lysed with urea, different solution digestion strategies were quantitatively assessed. In comparison with traditional proteolysis at 37 °C, urea in-solution digestion performed at room temperature improved peptide and protein identification and quantitation and had a minimum impact on miscleavage rates. Furthermore, the signal intensities and the number of carbamylated and pyroglutamic acid-modified peptides decreased. Overall, this led to a reduction in the negative effects often observed for such modifications. Data are available via ProteomeXchange with identifier PXD009426.},
  author       = {Betancourt, Lazaro and Sanchez, Aniel and Pla Parada, Indira and Kuras, Magdalena and Zhou, Qimin and Andersson, Roland and Marko-Varga, György},
  issn         = {1535-3893},
  keyword      = {Urea,proteomics},
  language     = {eng},
  month        = {05},
  number       = {7},
  pages        = {2556--2561},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {Quantitative Assessment of Urea In-Solution Lys-C/Trypsin Digestions Reveals Superior Performance at Room Temperature over Traditional Proteolysis at 37 °C.},
  url          = {http://dx.doi.org/10.1021/acs.jproteome.8b00228},
  volume       = {17},
  year         = {2018},
}