Analysis of Murine Natural Killer Cell Microsomal Proteins using Two Dimensional Liquid Chromatography Coupled to Tandem Electrospray Ionization Mass Spectrometry

BLONDER J,; RODRIGUEZ GALAN MC; CHAN KC; LUCAS AD; YU LR; CONRADS TP; ISSAQ HJ; YOUNG HA; VEENSTRA TD

AMER CHEMICAL SOC

Año: 2004 vol. 3 p. 862 - 862

his study describes the application of a single tube sample preparation technique coupled withmultidimensional fractionation for the analysis of a complex membrane protein sample from murinenatural killer (NK) cells. A solution-based method that facilitates the solubilization and tryptic digestion of integral membrane proteins is conjoined with strong cation exchange (SCX) liquid chromatography (LC) fractionation followed by microcapillary reversed-phase (íRP) LC tandem mass spectrometric analysis of each SCXLC fraction in second dimension. Sonication in buffered methanol solution was employed to solubilize, and tryptically digest murine NK cell microsomal proteins, allowing for the large-scale identification of integral membrane proteins, including the mapping of the membranespanning peptides. Bioinformatic analysis of the acquired tandem mass spectra versus the murine genome database resulted in 11 967 matching tryptic peptide sequences, corresponding to 5782 unique pept