Gas Phase Ion Chemistry to Determine Isoaspartate in a Peptide Backbone
Proof of concept evidence is presented for a new method for the determination of isoaspartate, an important post-translational modification. Chemical derivatization is performed using common reagents for the modification of carboxylic acids and shown to yield suitable diagnostic information with regard to isomerization at the aspartate residue. The diagnostic gas phase chemistry is probed by collision-induced dissociation mass spectrometry, on the timescale of the MS experiment and semi-quantitative calibration of the percentage of isoaspartate in a peptide sample is demonstrated.
KeywordsMass spectrometry Isobaric peptide Post-translational modification Nanoelectrospray ionization
The authors acknowledge financial support of this work by Amgen Inc. and appreciate valuable discussions with Dalton Snyder and Izydor Apostol.
- 3.Alfaro, J.F., Gillies, L.A., Sun, H.G., Dai, S., Zhang, T., Klaene, J., Kim, B.J., Lowenson, J.D., Clarke, S.G., Karger, B.L., Zhou, Z.S.: Chemo-enzymatic detection of protein isoaspartate using protein isoaspartate methyltransferase and hydrazine trapping. Anal. Chem. 80, 3882–3889 (2008)CrossRefPubMedGoogle Scholar
- 6.Hao, P., Qian, J., Dutta, B., Cheow, E.S., Sim, K.H., Meng, W., Adav, S.S., Alpert, A., Sze, S.K.: Enhanced separation and characterization of deamidated peptides with RP-ERLIC-based multidimensional chromatography coupled with tandem mass spectrometry. J. Proteome Res. 11, 1804–1811 (2012)CrossRefPubMedGoogle Scholar
- 7.Seebach, D., Gardiner, J.: b-Peptides: a surprise at every turn. Chem. Comm. 0, 2015–2022 (1997)Google Scholar
- 11.Zheng, X., Deng, L., Baker, E.S., Ibrahim, Y.M., Petyuk, V.A., Smith, R.D.: Distinguishing d- and l-aspartic and isoaspartic acids in amyloid beta peptides with ultrahigh resolution ion mobility spectrometry. Chem. Commun. (Camb). (2017)Google Scholar