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The analysis of multiple O-phosphoseryl-containing peptides by fast atom bombardment mass spectrometry

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Summary

Positive and negative ion FAB mass spectrometry were found to be useful for the structural analysis of phosphorylated peptides containing multiple O-phosphoseryl residues. The positive ion FAB mass spectra obtained for Ac-Ser(P)-Ser(P)-NHMe and Ac-Ser(P)-Ser(P)-Ser(P)-NHMe showed that β-eliminative loss of H3PO4 from the Ser(P)-residue was a major event in the fragmentation of the two phosphopeptides and that successive losses of H3PO4 from the [M+H]+ ion occurred when the Ser(P)-cluster was located at the N-terminus. In contrast, the FAB mass spectrum of Ac-Glu-Ser(P)-Leu-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-NHMe showed only a single loss of H3PO4 from the [M+H]+ ion, with further losses of H3PO4 from internal Ser(P)-residues only occurring when fragmentation of the parent phosphopeptide generated daughter fragments that contained (part of) an N-terminal Ser(P)-residue. Negative ion FAB mass spectrometry also proved useful for the structural analysis of the three Ser(P)-peptides and showed high-intensity [M-H]- ions along with minor [M-H-80]- fragment ions.

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Abbreviations

Ac:

acetyl

ΔAla:

dehydroalanyl

FAB-MS:

fast atom bombardment mass spectrometry

LSIMS:

liquid secondary ion mass spectrometry

NHMe:

N-methylamide

Ser(P):

O-phosphoseryl

Thr(P):

O-phosphothreonyl

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Author notes

  1. Alan L. Chaffee

    Present address: BHP Research, Melbourne Laboratories, 3170, Mulgrave, Victoria, Australia

Authors and Affiliations

  1. School of Chemistry, The University of Melbourne, 3052, Parkville, Victoria, Australia

    J. W. Perich & R. B. Johns

  2. Division of Fuel Chemistry, CSIRO, 2234, Menai, New South Wales, Australia

    Imantz Liepa & Alan L. Chaffee

Authors
  1. J. W. Perich
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  2. Imantz Liepa
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  3. Alan L. Chaffee
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  4. R. B. Johns
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Perich, J.W., Liepa, I., Chaffee, A.L. et al. The analysis of multiple O-phosphoseryl-containing peptides by fast atom bombardment mass spectrometry. Lett Pept Sci 2, 345–351 (1996). https://doi.org/10.1007/BF00119998

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  • DOI: https://doi.org/10.1007/BF00119998

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