Proteomic investigation of natural killer cells microsomas using gas-phase fractionation by mass spectrometry.

BLONDER J; RODRIGUEZ GALAN MC; LUCAS DA; YOUNG HA; ISAAQ HJ; VEENSTRA TD; CONRADS TP

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Año: 2004 vol. 1698 p. 87 - 87

We have explored the utility of gas-phase fractionation by mass spectrometry (MS) in the mass-to-charge (m/z) dimension (GPFm/z) for increasing the effective number of protein identifications in cases where sample quantity limits the use of multi-dimensional chromatographicfractionation. 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 GPFm/z using eight narrow precursor ion scan m/z ranges enabled the identification of 340 NK cell proteins from 12 Ag 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 GPFm/z represents an effective technique for increasing protein identifications in global proteomic investigations especial