title: Phosphopeptide analysis by directly coupling two-dimensional planar electrochromatography/thin-layer chromatography with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
abstract: A novel strategy is presented for the fractionation of complex peptide mixtures using two-dimensional planar electrochromatography/thin-layer chromatography (2D PEC/TLC). Phosphopeptides migrate more slowly in the first dimension, based upon their anionic phosphate residues, and certain predominantly acidic phosphopeptides even migrate in the opposite direction, relative to the bulk of the peptides. Phosphopeptides are further distinguished based upon hydrophilicity in the second dimension. This permits a restricted region of the plate to be directly interrogated for the presence of phosphopeptides by mass spectrometry (MS). Phosphopeptide analysis from the plates by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF)-MS and tandem MS enabled peptide sequencing and identification.
authors.affiliation: Biochemistry Department, PerkinElmer Life and Analytical Sciences, Waltham, MA 02451, USA.
authors.forename: Venkateswarlu
authors.initials: V
authors.lastname: Panchagnula
authors.forename: Alvydas
authors.initials: A
authors.lastname: Mikulskis
authors.forename: Linan
authors.initials: L
authors.lastname: Song
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.forename: Mei
authors.initials: M
authors.lastname: Wang
authors.forename: Tanya
authors.initials: T
authors.lastname: Knubovets
authors.forename: Elaine
authors.initials: E
authors.lastname: Scrivener
authors.forename: Eva
authors.initials: E
authors.lastname: Golenko
authors.forename: Ira S
authors.initials: IS
authors.lastname: Krull
authors.forename: Michael
authors.initials: M
authors.lastname: Schulz
authors.lastname: Heinz-Emil-Hauck
authors.forename: Wayne F
authors.initials: WF
authors.lastname: Patton
mesh_terms: D053801:Capillary Electrochromatography / Q000379:methods*; D002855:Chromatography, Thin Layer / Q000379:methods*; D010455:Peptides / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D010750:Phosphoproteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
chemical_list: D010455:Peptides; D010750:Phosphoproteins; D014357:Trypsin