Proteomic Investigation of Natural Killer Cell Microsomes Using Gas-Phase Fractionation by Mass Spectrometry
Biochimica et Biophysica Acta
We have explored the utility of gas-phase fractionation by mass spectrometry (MS) in the mass-to-charge (m/z) dimension (GPF(m/z)) for increasing the effective number of protein identifications in cases where sample quantity limits the use of multi-dimensional chromatographic fractionation. A peptide digestate from proteins isolated from the membrane fraction of natural killer (NK) cells was analyzed by microcapillary reversed-phase liquid chromatography coupled online to an ion-trap (IT) mass spectrometer. Performing GPF(m/z) using eight narrow precursor ion scan m/z ranges enabled the identification of 340 NK cell proteins from 12 microg of digestate, representing more than a fivefold increase in the number of proteins identified as compared to the same experiment employing a standard precursor ion survey scan m/z range (i.e., m/z 400-2000). The results show that GPF(m/z) represents an effective technique for increasing protein identifications in global proteomic investigations especially when sample quantity is limited.
Amino acid sequence, cation transport proteins, killer cells, natural, mass spectrometry, membrane proteins, microsomes, peptides, proteome
Blonder, Josip; Rodriguez-Galan, Maria Cecilia; Lucas, David A.; Young, Howard A.; Issaq, Haleem J.; Veenstra, Timothy D.; and Conrads, Thomas P., "Proteomic Investigation of Natural Killer Cell Microsomes Using Gas-Phase Fractionation by Mass Spectrometry" (2004). Pharmaceutical Sciences Faculty Publications. 481.