November 16, 2007
Journal Article

Phosphopeptide elution times in reversed-phase liquid chromatography

Abstract

Elution time shifts between 33 different peptides and their corresponding phosphopeptides ranging from 4 amino acid residues to 35 amino acids in length were systematically investigated utilizing a high resolution reversed-phase liquid chromatography (RPLC) system. Observed peptide elution time shifts for a single phosphorylation ranged from -5.28 min (for pYVPML) to +0.59 min (for HRDpSGLLDSLGR). Peptides containing a phosphotyrosine residue displayed a significant decrease in elution time following phosphorylation compared to their similar-sized peptides with phosphoserine or phosphothreonine residues. While the observed elution time generally decreased due to phosphorylation, five peptides displayed increased elution time as a result of phosphorylation. For large peptides (= 18 amino acids), the elution time shifts due to single phosphorylation were limited (ranging between -0.48 min and +0.03 min), while the elution time shifts for small peptides (

Revised: October 12, 2015 | Published: November 16, 2007

Citation

Kim J., K. Petritis, Y. Shen, D.G. Camp, R.J. Moore, and R.D. Smith. 2007. "Phosphopeptide elution times in reversed-phase liquid chromatography." Journal of Chromatography A 1172, no. 1:9-18. PNNL-SA-53249. doi:doi:10.1016/j.chroma.2007.09.032