title: Selective isolation-detection of two different positively charged peptides groups by strong cation exchange chromatography and matrix-assisted laser desorption/ionization mass spectrometry: application to proteomics studies.
abstract: We report here a procedure for the independent analysis of two groups of peptides by liquid chromatography-matrix-assisted laser desorption/ionization mass spectrometry (LC-MALDI MS/MS), using a selective isolation-detection procedure. In this procedure all primary amino groups of tryptic peptides derived from mouse liver proteins are blocked, restricting their positive charge, at acidic pH, to the presence of histidine and arginine residues. After strong cation exchange chromatography, multiply charged peptides (R + H > 1) are retained on the column and separated with high selectivity from singly (R + H = 1) and neutral peptides (R + H = 0) which are together collected in the flow-through. Using LC-MALDI-MS/MS analysis, the retained fraction displayed a 94% of enrichment of multiply charged peptides while in the flow-through; peptides with at least one arginine or histidine residue were exclusively identified, which suggests that MS detection in this fraction is restricted only to those peptides with ionizable side chains, arginine and histidine amino acids.
authors.forename: Aniel
authors.initials: A
authors.lastname: Sanchez
authors.forename: Wei
authors.initials: W
authors.lastname: Sun
authors.forename: Lan
authors.initials: L
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Ma
authors.forename: Lazaro
authors.initials: L
authors.lastname: Betancourt
authors.forename: Jeovanis
authors.initials: J
authors.lastname: Gil
authors.forename: Yasset
authors.initials: Y
authors.lastname: Perez-Riverol
authors.forename: Jorge
authors.initials: J
authors.lastname: Fernandez de-Cossio
authors.forename: Gabriel
authors.initials: G
authors.lastname: Padron
authors.forename: Ying
authors.initials: Y
authors.lastname: Jiang
authors.forename: Fuchu
authors.initials: F
authors.lastname: He
authors.forename: Luis Javier
authors.initials: LJ
authors.lastname: Gonzalez
authors.forename: Vladimir
authors.initials: V
authors.lastname: Besada
mesh_terms: D000595:Amino Acid Sequence; D002412:Cations; D002852:Chromatography, Ion Exchange / Q000379:methods*; D007477:Ions; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D002412:Cations; D007477:Ions; D010455:Peptides