title: Selective isolation of multiply charged peptides: a confident strategy for protein identification using a linear trap quadrupole mass spectrometer.
abstract: In this work, a method devised for the selective isolation of multiply-charged peptide applied to a complex protein mixture was evaluated for the first time using a mass spectrometer with low resolution (LTQ). In this procedure, all primary amino groups of tryptic peptides derived from human Liver tissue interstitial fluid (TIF) 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 in the column and separated with high selectivity from singly (#R+#H = 1) and neutral peptides (#R +#H = 0) which are collected together in the flow-through. Using Liquid chromatography electrospray ionization tandem mass spectrometry analysis the retained fraction displayed a 95% enrichment of multiply charged peptides while in the flow-through; only 4% of multiply-charged peptides were identified.
authors: Department of Proteomics and Bioinformatics. Center for Genetic Engineering and Biotechnotogy, Havana, Cuba.
authors: Aniel
authors: A
authors: Sanchez
authors: Wei
authors: W
authors: Sun
authors: Jie
authors: J
authors: Ma
authors: Lazaro
authors: L
authors: Betancourt
authors: Yasset
authors: Y
authors: Perez-Riverol
authors: Jorge Fernandez
authors: JF
authors: de-Cossio
authors: Gabriel
authors: G
authors: Padron
authors: Ying
authors: Y
authors: Jiang
authors: Fuchu
authors: F
authors: He
authors: Luis Javier
authors: LJ
authors: Gonzalez
authors: Vladimir
authors: V
authors: Besada
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D045604:Extracellular Fluid / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry; D006801:Humans; D008099:Liver / Q000737:chemistry*; D008239:Lysine / Q000737:chemistry; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification*; D020543:Proteome / Q000737:chemistry*; D040901:Proteomics / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D010455:Peptides; D020543:Proteome; D006639:Histidine; D008239:Lysine