title: Predicting Electrophoretic Mobility of Tryptic Peptides for High-Throughput CZE-MS Analysis.
abstract: A multiparametric sequence-specific model for predicting peptide electrophoretic mobility has been developed using large-scale bottom-up proteomic CE-MS data (5% (∼0.8M) acetic acid as background electrolyte). Peptide charge (Z) and size (molecular mass, M) are the two major factors determining electrophoretic mobility, in complete agreement with previous studies. The extended size of the data set (>4000 peptides) permits access to many sequence-specific factors that impact peptide mobility. The presence of acidic residues Asp and Glu near the peptide N-terminus is by far the most prominent among them. The induction effect of the side chain of N-terminal Asp reduces the basicity of the N-terminal amino group and, as hence, its charge, by ∼0.27 units, lowering mobility. The correlation of the model (R2∼ 0.995) indicates that the peptide separation process in CZE is relatively simple and can be predicted to a much higher precision than current RP-HPLC models. Similar to RP-HPLC prediction studies, we anticipate future developments that introduce peptide migration standards, collect larger data sets for modeling through the alignment of multiple CZE-MS acquisitions, and study of the behavior of peptides carrying post-translational modifications. The increased size of data sets will also permit investigation of the fine-scale effects of peptide secondary structure on peptide mobility. We observed that peptides with higher helical propensity tend to have higher than predicted electrophoretic mobility; the incorporation of these features into CZE migration models will require significantly larger data sets.
authors.forename: Oleg V
authors.identifier: 0000-0002-9989-6593
authors.initials: OV
authors.lastname: Krokhin
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Anderson
authors.forename: Vic
authors.initials: V
authors.lastname: Spicer
authors.affiliation: Department of Chemistry, Michigan State University , East Lansing, Michigan 48824, United States.
authors.forename: Liangliang
authors.initials: L
authors.lastname: Sun
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame , Notre Dame, Indiana 46556, United States.
authors.forename: Norman J
authors.initials: NJ
authors.lastname: Dovichi
mesh_terms: D001224:Aspartic Acid / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D019075:Electrophoresis, Capillary / Q000379:methods*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D018698:Glutamic Acid / Q000032:analysis; D006367:HeLa Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*; D014357:Trypsin / Q000737:chemistry*
chemical_list: D010455:Peptides; D001224:Aspartic Acid; D018698:Glutamic Acid; D014357:Trypsin