Quantitative Assessment of Urea In-Solution Lys-C/Trypsin Digestions Reveals Superior Performance at Room Temperature over Traditional Proteolysis at 37 °C.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1fdd9bb8-afc7-47c9-8519-21ee5f6fb7f6
- author
- Betancourt, Lazaro LU ; Sanchez, Aniel LU ; Pla Parada, Indira LU ; Kuras, Magdalena LU ; Zhou, Qimin LU ; Andersson, Roland LU and Marko-Varga, György LU
- organization
-
- Clinical Protein Science and Imaging (research group)
- Department of Clinical Sciences, Lund
- Clinical Chemistry, Malmö (research group)
- BioCARE: Biomarkers in Cancer Medicine improving Health Care, Education and Innovation
- Hepato-Pancreato-Biliary Surgery (research group)
- Surgery (Lund)
- CEBMMS PI (research group)
- Mass Spectrometry
- Department of Biomedical Engineering
- publishing date
- 2018-05-29
- 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 (ACS)
- external identifiers
-
- scopus:85047969180
- pmid:29812944
- ISSN
- 1535-3893
- DOI
- 10.1021/acs.jproteome.8b00228
- language
- English
- LU publication?
- yes
- id
- 1fdd9bb8-afc7-47c9-8519-21ee5f6fb7f6
- alternative location
- https://www.scopus.com/inward/record.uri?eid=2-s2.0-85047969180&doi=10.1021%2facs.jproteome.8b00228&partnerID=40&md5=c77937b541399d7615e0b3e3a8a08948
- date added to LUP
- 2018-12-13 15:07:46
- date last changed
- 2022-03-17 19:55:50
@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}}, keywords = {{Urea; proteomics}}, language = {{eng}}, month = {{05}}, number = {{7}}, pages = {{2556--2561}}, publisher = {{The American Chemical Society (ACS)}}, 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}}, doi = {{10.1021/acs.jproteome.8b00228}}, volume = {{17}}, year = {{2018}}, }