The DOI matches its Crossref record; 24 of the 24 references it cites were found in a registry or an index.
Phosphopeptide Analysis by Matrix-Assisted Laser Desorption Time-of-Flight Mass Spectrometry
Abstract
In this paper we present methods for identifying and sequencing phosphopeptides in simple mixtures, such as HPLC fractions, at the subpicomole level by (+) ion matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-MS). Data are presented which indicate that when a reflectron time-of-flight mass spectrometer is used, MALDI can distinguish tyrosine phosphorylation from serine and threonine phosphorylation for peptides containing a single phosphate group. Phosphopeptides are identified in the (+) ion MALDI reflector spectrum by the presence of [MH-H3PO4]+ and [MH-HPO3]+ fragment ions formed by metastable decomposition. An abundant [MH-H3PO4]+ ion, accompanied by a weaker [MH-HPO3]+ ion indicates that the peptide is most likely phosphorylated on serine or threonine. In contrast, phosphotyrosine-containing peptides generally exhibit [MH-HPO3]+ fragment ions and little, if any [MH-H3PO4]+. Ambiguities do arise, most often with phosphopeptides that contain residues which readily lose water (such as unmodified serine), but these can often be resolved by recording a complete metastable fragment ion (postsource decay) spectrum. Postsource decay is shown here to be a viable technique for sequencing phosphopeptides. It can be used to distinguish between serine/ threonine and tyrosine phosphorylation and in many cases can be used to determine the exact site of phosphorylation in a peptide sequence. Nearly complete sequence coverage and phosphorylation site mapping is generally possible using approximately 300 fmol of peptide.
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Cites
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Cited by
Mass Spectrometry in Proteomics
Aebersold, R., & Goodlett, D. R. (2001) · Chemical Reviews · doi:10.1021/cr990076h · Cited by 1,238
Immobilized Gallium(III) Affinity Chromatography of Phosphopeptides
Posewitz, M. C., & Tempst, P. (1999) · Analytical Chemistry · doi:10.1021/ac981409y · Cited by 832
Tanner, K. G., Landry, J., Sternglanz, R., & Denu, J. M. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.250422697 · Cited by 555
Zhao, J., Du, Y., Horton, J. R., Upadhyay, A. K., Lou, B., Bai, Y., Zhang, X., Du, L., Li, M., Wang, B., et al. (2011) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1100012108 · Cited by 105
Lim, A., Prokaeva, T., McComb, M. E., O'Connor, P. B., Théberge, R., Connors, L. H., Skinner, M., & Costello, C. E. (2002) · Analytical Chemistry · doi:10.1021/ac010780+ · Cited by 63
Metadata
- Authors
- Annan, R. S., & Carr, S. A.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1996-10-01
- Volume
- 68
- Issue
- 19
- Pages
- 3413-3421
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W2071106732
- Cited by
- 310
- References
- 24
- FWCI
- 9.93
- Type
- journal article
- Language
- en
- Open access
- closed
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