Izvorni znanstveni članak

https://doi.org/10.5599/admet.520

Peptide retention time prediction for immobilized artificial membrane phosphatidylcholine stationary phase: method development and preliminary observations

Daniel Gussakovsky ; Department of Chemistry, University of Manitoba, 360 Parker Building, Winnipeg, Manitoba R3T 2N2, Canada
Haley Neustaeter ; Department of Chemistry, University of Manitoba, 360 Parker Building, Winnipeg, Manitoba R3T 2N2, Canada
Victor Spicer ; Manitoba Centre for Proteomics and Systems Biology, University of Manitoba, 799 JBRC, 715 McDermot Avenue, Winnipeg, Manitoba R3E 3P4, Canada
Oleg V. Krokhin ; Manitoba Centre for Proteomics and Systems Biology, University of Manitoba, 799 JBRC, 715 McDermot Avenue, Winnipeg, Manitoba R3E 3P4, Canada

Sažetak

Development of the first peptide retention prediction model for immobilized artificial membrane phosphatidylcholine (IAM.PC) stationary phase is reported. 2D liquid chromatography coupled to tandem mass spectrometry (2D LC-MS/MS) analysis of a whole cell lysate of S. cerevisiae yielded a retention dataset of ~29,500 tryptic peptides; sufficient for confident assignment of retention coefficients which determine the contribution of individual amino acids in peptide retention. Retention data from the first dimension was used for the modelling: an IAM.PC.DD2 column, with pH 7.4 ammonium bicarbonate, and a water/acetonitrile gradient. Peptide separation using the IAM.PC.DD2 phase was compared to a standard C18 phase (Luna C18(2)). There was a significant reduction in peptide retention (~14 % acetonitrile on average), indicating that the phosphatidylcholine stationary phase is significantly more hydrophilic. In comparison to the C18 phase, a substantial increase was found in the relative retention contribution for the positively charged Arg and Lys, and the aromatic Tyr, Trp and His residues. A decrease in retention contribution was observed for the negatively charged Asp and Glu. This indicates an involvement of electrostatic interactions with the glycerophosphate functional groups, and possibly, delocalization effects from hydrogen bonds between the phosphate group and the aromatic side chains in the separation mechanism.

Ključne riječi

Peptide retention modelling; immobilized artificial membrane chromatography

Hrčak ID:

201573

URI

https://hrcak.srce.hr/201573

Datum izdavanja:

16.6.2018.

Posjeta: 1.002 *