A General Strategy for the use of Mass Spectrometric Molecular Weight Information in Protein Purification and Sequence Determination

Abstract

At the MPSA 1988 in Berlin we presented a general strategy for the combined use of mass spectrometric molecular weight information and automatic Edman degradation for sequence determination of proteins (Roepstorff et al., 1989). Among the mass spectrometric methods available then, i.e. plasma desorption mass spectrometry (PDMS) and fast atom bombardment mass spectrometry (FABMS), we had chosen PDMS because this method in our experience yielded better sensitivity and higher practical mass range than FABMS. The mass accuracy although not as good as for FABMS was in most cases adequate. PDMS had further advantages over FABMS in simplicity in sample preparation, instrument operation and data interpretation. This has been of utmost importance for our routine use of mass spectrometry in protein sequencing. In the past years we have sequenced several proteins using this strategy (Andersen et al., in press; Andreassen et al., submitted; Hojrup et al., 1986; 1991; Klarskov et al., 1989; Mikkelsen et al., 1989; Parello et al., in preparation; Talbo et al., 1990).