Abstract
Electron-transfer dissociation (ETD) with supplemental activation of the doubly charged deamidated tryptic digested peptide ions allows differentiation of isoaspartic acid and aspartic acid residues using the c + 57 or z • − 57 peaks. The diagnostic peak clearly localizes and characterizes the isoaspartic acid residue. Supplemental activation in ETD of the doubly charged peptide ions involves resonant excitation of the charge reduced precursor radical cations and leads to further dissociation, including extra backbone cleavages and secondary fragmentation. Supplemental activation is essential to obtain a high quality ETD spectrum (especially for doubly charged peptide ions) with sequence information. Unfortunately, the low-resolution of the ion trap mass spectrometer makes detection of the diagnostic peak, [M-60], for the aspartic acid residue difficult due to interference with side-chain loss from arginine and glutamic acid residues.
Article PDF
Avoid common mistakes on your manuscript.
References
Robinson, N. E.; Robinson, A. B. Molecular Clocks: Deamidation of Asparaginyl and Glutaminyl Residues in Peptides and Proteins; Althouse Press: Cave Junction, OR, 2004.
Clarke, S. Propensity for Spontaneous Succinimide Formation from Aspartyl and Asparaginyl Residues in Cellular Proteins. Int. J. Pept. Protein Res. 1987, 30, 808–821.
Radkiewics, J. L.; Zipse, H.; Clarke, S.; Houk, K. N. Accelerated Racemization of Aspartic Acid and Asparagine Residues Via Succinimide Intermediates: An Ab Initio Theoretical Exploration of Mechanism. J. Am. Chem. Soc. 1996, 118, 9148–9155.
Geiger, T.; Clarke, S. Deamidation, Isomerization, and Racemization at Asparaginyl and Aspartyl Residues in Peptides. J. Biol. Chem. 1987, 262, 785–795.
Robinson, N. E.; Robinson, A. B. Molecular Clocks. Proc. Natl. Acad. Sci. U.S.A. 2001, 98, 944–949.
Robinson, N. E.; Robinson, A. B. Prediction of Protein Deamidation Rates from Primary and Three-Dimensional Structure. Proc. Natl. Acad. Sci. U.S.A. 2001, 98, 4367–4372.
Roher, A. E.; Lowenson, J. D.; Clarke, S.; Wolkow, C.; Wang, R.; Cotter, R. J.; Reardon, I. M.; Zurcher-Neely, H. A.; Heinrikson, R. L.; Ball, M. J. Structural Alterations in the Peptide Backbone of β-Amyloid Core Protein may account for its Deposition and Stability in Alzheimer’s Disease. J. Biol. Chem. 1993, 268, 3072–3083.
Shimizu, T.; Watanabe, A.; Ogawara, M.; Mori, H.; Shirasawa, T. Isoaspartate Formation and Neurodegeneration in Alzheimer’s Disease. Arch. Biochem. Biophys. 2000, 381, 225–234.
Sargaeva, N. P.; Lin, C.; O’Connor, P. B. Identification of Aspartic and Isoaspartic Acid Residues in Amyloid β Peptides, Including Aβ1-42, Using Electron-Ion Reactions. Anal. Chem. 2009, 81, 9778–9786.
Hsu, Y. R.; Chang, W. C.; Mendiza, E. A.; Hara, S.; Chow, D. T.; Mann, M. B.; Langley, K. E.; Lu, H. S. Selective Deamidation of Recombinant Human Stem Cell Factor During In Vitro Aging; Isolation, and Characterization of the Aspartyl and Isoaspartyl Homodimers and Heterodimers. Biochemistry 1998, 37, 2251–2262.
Lehmann, W.; Schlosser, A.; Erben, G.; Pipkorn, R.; Bossemeyer, D.; Kinzel, V. Analysis of Isoaspartate in Peptides by Electrospray Tandem Mass Spectrometry. Protein Sci. 2000, 9, 2260–2268.
Schindler, P.; Muller, D.; Marki, W.; Grossenbacher, H.; Richter, W. J. Characterization of a β-Asp33 Isoform of Recombinant Hirudin Sequence Variant 1 by Low-Energy Collision-Induced Dissociation. J. Mass Spectrom. 1996, 31, 967–974.
Cournoyer, J. J.; Pittman, J. L.; Ivlevaver, A. B.; Fallows, E.; Waskell, L.; Costello, C. E.; O’Connor, P. B. Deamidation: Differentiation of Aspartyl from Isoaspartyl Products in Peptides by Electron Capture Dissociation. Protein. Sci. 2005, 14, 452–463.
Cournoyer, J. J.; Lin, C.; O’Connor, P. B. Detecting Deamidation Products in Proteins by Electron Capture Dissociation. Anal. Chem. 2006, 78, 1264–1271.
Cournoyer, J. J.; Lin, C.; Bowman, M. J.; O’Connor, P. B. Quantitating the Relative Abundance of Isoaspartyl Residues in Deamidated Proteins by Electron Capture Dissociation. J. Am. Soc. Mass Spectrom. 2007, 18, 48–56.
Li, X. J.; Cournoyer, J. J.; Lin, C.; O’Connor, P. B. Use of 18O Labels to Monitor Deamidation during Protein and Peptide Sample Processing. J. Am. Soc. Mass Spectrom. 2008, 19, 855–864.
O’Connor, P. B.; Cournoyer, J. J.; Pitteri, S. J.; Chrisman, P. A.; McLuckey, S. A. Differentiation of Aspartic and Isoaspartic Acids Using Electron Transfer Dissociation. J. Am. Soc. Mass Spectrom. 2006, 17, 15–19.
Andreazza, H. J.; Wang, T. F.; Bagley, C. J.; Hoffmann, P.; Bowie, J. H. Negative Ion Fragmentations of Deprotonated Peptides. The Unusual Case of isoAsp: A Joint Experimental and Theoretical Study. Comparison with Positive Ion Cleavages. Rapid Commun. Mass Spectrom. 2009, 23, 1993–2002.
Pitteri, S. J.; Chrisman, P. A.; Hogan, J. M.; McLuckey, S. A. Electron Transfer Ion/Ion Reactions in a Three-Dimensional Quadrupole Ion Trap: Reactions of Doubly and Triply Protonated Peptides with SO2. Anal. Chem. 2005, 77, 1831–1839.
Swaney, D. L.; McAlister, G. C.; Wirtala, M.; Schwartz, J. C.; Syka, J. E. P.; Coon, J. J. Supplemental Activation Method for High-Efficiency Electron-Transfer Dissociation of Doubly Protonated Peptide Precursors. Anal. Chem. 2007, 79, 477–485.
Domon, B.; Bodenmiller, B.; Carapito, C.; Hao, Z. Q.; Huehmer, A.; Aebersold, R. Electron Transfer Dissociation in Conjunction with Collision Activation to Investigate the Drosophila melanogaster Phosphoproteome. J. Proteome Res. 2008, 8, 2633–2639.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Chan, W.Y.K., Chan, T.W.D. & O’Connor, P.B. Electron transfer dissociation with supplemental activation to differentiate aspartic and isoaspartic residues in doubly charged peptide cations. J Am Soc Mass Spectrom 21, 1012–1015 (2010). https://doi.org/10.1016/j.jasms.2010.02.002
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1016/j.jasms.2010.02.002