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Application of a Combined Weak Cation-Exchange/Crown Ether Column: First Demonstrations of a Versatile Tool for Proteome Subselection

Robin Tuytten, Bart Ruttens, Katelijne Gheysen UGent, Koen Sandra, Koen De Cremer, Dominique Vlieghe, Natalie Van Landuyt, Gregoire Thomas, José Martins UGent and Patrick Sandra UGent, et al. (2009) ANALYTICAL CHEMISTRY. 81(7). p.2456-2469
abstract
The present paper introduces the use of a weak cation-exchange/crown ether column in the proteomics field. The 18-crown-6 ether functionality is well-known to selectively complex ammonium and monoalkylammonium, ions, which should make this column highly suitable to trap peptides with free alpha-NH2 or free epsilon-NH2 groups from lysine side chains. This unique selection mechanism was put to the test in an N-teromics setup which aims for the enrichment of deliberately acetylated protein N-terminal peptides from a serum digest. It was demonstrated that peptides with free alpha-NH2 groups and peptides with alpha-amino-acetylated groups can be separated from each other using this weak cation-exchange/crown ether column. The peptides of interest, bearing no free primary amines, were found to be significantly enriched in the column's flow through. At the same time a favorable coenrichment of N-glycosylated peptides was observed. To obtain more insight in the contributions of the two distinct column functionalities, i.e., the weak cation exchanger and the crown ether, the experimental data were checked against a theoretical prediction of the outcome.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
journal title
ANALYTICAL CHEMISTRY
Anal. Chem.
volume
81
issue
7
pages
2456 - 2469
Web of Science type
Article
Web of Science id
000264759400009
JCR category
CHEMISTRY, ANALYTICAL
JCR impact factor
5.214 (2009)
JCR rank
3/70 (2009)
JCR quartile
1 (2009)
ISSN
0003-2700
DOI
10.1021/ac801975b
language
English
UGent publication?
yes
classification
A1
id
622509
handle
http://hdl.handle.net/1854/LU-622509
date created
2009-05-11 11:15:32
date last changed
2013-01-30 09:41:51
@article{622509,
  abstract     = {The present paper introduces the use of a weak cation-exchange/crown ether column in the proteomics field. The 18-crown-6 ether functionality is well-known to selectively complex ammonium and monoalkylammonium, ions, which should make this column highly suitable to trap peptides with free alpha-NH2 or free epsilon-NH2 groups from lysine side chains. This unique selection mechanism was put to the test in an N-teromics setup which aims for the enrichment of deliberately acetylated protein N-terminal peptides from a serum digest. It was demonstrated that peptides with free alpha-NH2 groups and peptides with alpha-amino-acetylated groups can be separated from each other using this weak cation-exchange/crown ether column. The peptides of interest, bearing no free primary amines, were found to be significantly enriched in the column's flow through. At the same time a favorable coenrichment of N-glycosylated peptides was observed. To obtain more insight in the contributions of the two distinct column functionalities, i.e., the weak cation exchanger and the crown ether, the experimental data were checked against a theoretical prediction of the outcome.},
  author       = {Tuytten, Robin and Ruttens, Bart and Gheysen, Katelijne and Sandra, Koen and De Cremer, Koen and Vlieghe, Dominique and Van Landuyt, Natalie and Thomas, Gregoire and Martins, Jos{\'e} and Sandra, Patrick and Kas, Koen and Verleyen, Katleen},
  issn         = {0003-2700},
  journal      = {ANALYTICAL CHEMISTRY},
  language     = {eng},
  number       = {7},
  pages        = {2456--2469},
  title        = {Application of a Combined Weak Cation-Exchange/Crown Ether Column: First Demonstrations of a Versatile Tool for Proteome Subselection},
  url          = {http://dx.doi.org/10.1021/ac801975b},
  volume       = {81},
  year         = {2009},
}

Chicago
Tuytten, Robin, Bart Ruttens, Katelijne Gheysen, Koen Sandra, Koen De Cremer, Dominique Vlieghe, Natalie Van Landuyt, et al. 2009. “Application of a Combined Weak Cation-Exchange/Crown Ether Column: First Demonstrations of a Versatile Tool for Proteome Subselection.” Analytical Chemistry 81 (7): 2456–2469.
APA
Tuytten, R., Ruttens, B., Gheysen, K., Sandra, K., De Cremer, K., Vlieghe, D., Van Landuyt, N., et al. (2009). Application of a Combined Weak Cation-Exchange/Crown Ether Column: First Demonstrations of a Versatile Tool for Proteome Subselection. ANALYTICAL CHEMISTRY, 81(7), 2456–2469.
Vancouver
1.
Tuytten R, Ruttens B, Gheysen K, Sandra K, De Cremer K, Vlieghe D, et al. Application of a Combined Weak Cation-Exchange/Crown Ether Column: First Demonstrations of a Versatile Tool for Proteome Subselection. ANALYTICAL CHEMISTRY. 2009;81(7):2456–69.
MLA
Tuytten, Robin, Bart Ruttens, Katelijne Gheysen, et al. “Application of a Combined Weak Cation-Exchange/Crown Ether Column: First Demonstrations of a Versatile Tool for Proteome Subselection.” ANALYTICAL CHEMISTRY 81.7 (2009): 2456–2469. Print.