Fast atom bombardment peptide sequencing

Kbnig, W.A. Aydin, M. Schulze, U. Rapp, U. Hbhn, M. Pesch, R. Kalik-hevitch, V.N. Fast-Atom-Bombardment for Peptide Sequencing - a Comparison With Conventional Ionization Techniques. Int. J. Mass Spectrom. Ion Phys. 1983, 46, 403-406. 

Fast atom bombardment mass spectrometry (FABMS) has become an important addition to the ionization techniques available to the analytical chemist in recent years. It has been particularly useful in a number of diverse applications which include molecular weight determinations at high mass, peptide and oligosaccharide sequencing, structural analysis of organic compounds, determination of salts and metal complexes, and the analysis of ionic species in aqueous solutions. This paper will focus on some aspects of the quantitative measurement of ionic species in solution. The reader is referred to a more comprehensive review for more details of some of the examples given here as well as other applications (1). 

Short polypeptides can be sequenced rapidly by fast atom bombardment mass spectrometry (FAB-MS). This technique not only provides the amino acid sequence of the peptide but also information on post-translational modifications. 

Fast atom bombardment 20,000 Ability to analyze mixtures MW and limited sequence information for pure small peptides adaptable for LC/MS (NP- and RP-HPLC) widely available Matrix interference below m/z 500 widely variable response peptide-induced suppression effects limited to flow rates 10 /xl/min... 

Finally, disulphide bonds can be located by hydrolysing a protein to a mixture of peptides using either a proteinase or a specific chemical method of cleavage and the mixture can be analysed directly by fast-atom bombardment mass spectrometry (Chapter 3) and again after reduction of disulphide bonds (Yazdanparast et al., 1987). By identifying those peaks which disappear as a result of reduction and new peaks with appropriate masses that have taken their place, it is simple to assign disulphide bonds to the relevant amino-acid sequences. 

The following ion-activation techniques have been used at one time or other to sequence peptides (1) fast atom bombardment (FAB) ionization, (2) CID—tandem MS (MS/MS), (3) ESI in-source CID, (4) MALDI ion-source decay, (5) MALDI postsource decay (PSD), (6) electron-capture dissociation (ECD) and electron-transfer dissociation, and (7) peptide ladder sequencing. Because of the lack of space, only (2) and (4) will be discussed further. 

In addition to molecvdar weight information. Fast Atom Bombardment Mass Spectrometry can also be used for sequencing polypeptides and oHgonucleotides. One such example is the study of the amino acid sequence in the ohgopeptide Antiamoebin I by Taylor et al. (30) by a combination of FAB and B/E-hnked scans. This peptide contains 16 amino acids. The peak at m/z 1670 is the quasi-molecular ion [M -t HF. A linked-scan experiment of the B/E type performed on this ion produced a peak at m/z 1422 which presented confirmatory evidence a loss of the terminal prohne.phenylalaninol.H group. A similar B/E scan from the latter ion produces ions at m/z 1337 and m/z 1244 for the loss of aminobutyric acid and hydroxyprohne.amino-isobutyric acid, respectively. This process can be repeated vmtil aU the amino acids are... 

Sequences of peptides may be analysed by the reduction of N-acetylated peptides to N-ethylpolyamino alcohols followed by interpretation of the mass spectra of the O-TMS derivatives [55, 56]. The latter have excellent mass spectrometric properties, but the GC-MS method has been largely supplanted by developments in direct mass spectrometric methods such as fast atom bombardment (see Chapter 12). 

Mass spectrometry (MS) in its different detection modes is now the ultimate method for the characterization of the mass, composition, and sequence of peptide or protein components in a mixture and is particularly well suited to the mobile phases employed in RPC. Electrospray ionization (ESI) and matrix-assisted laser desorption ionization time-of-fiight (MALDl-TOF) mass spectroscopic methods, and to a lesser extent thermospray and fast atom bombardment... 

Morris HR, Panico M, Barber M, Bordoli RS, Sedgwick RD, Tyler A. Fast atom bombardment a new mass spectrometric method for peptide sequence analysis. Biochem Biophys Res Commiin. 1981 101 623-31. 

A. N. Fast Atom Bombardment a New Mass Spectrometric Method for Peptide Sequence Analysis. Biochem. Biophys. Res. Commun. 1981,707,623-631. 

Ishikawa, K. and Niva, Y, Computer-aided peptide sequencing by fast atom bombardment mass spectrometry. Biomed. Environ. Mass Spectrom., 13, 373-380 (1986). 

Bradley, C. V., D. H. Williams, and M. R. Hanley Peptide Sequencing Using the Combination of Edman Degradation, Carboxypeptidase Digestion and Fast Atom Bombardment Mass Spectrometry. Biochem. Biophys. Res. Comm. 104, 1223 (1982). Williams, D. H., S. Santikarn, P. B. Oelrichs, F. de Angelis, J. K. Macleod, and R. J. Smith The Structure of a Toxic Octapeptide, Containing 4 D-amino acids, from the Larvae of a Sawfly. Lophyrotoma Interrupta. J. Chem. Soc. (London), Chem. Commun. 1982, 1394. 

---