{"id":816,"date":"2012-01-05T13:28:50","date_gmt":"2012-01-05T18:28:50","guid":{"rendered":"https:\/\/www.bumc.bu.edu\/ftms\/?page_id=816"},"modified":"2016-09-09T15:55:42","modified_gmt":"2016-09-09T19:55:42","slug":"isoaspartome","status":"publish","type":"page","link":"https:\/\/www.bumc.bu.edu\/ftms\/research\/isoaspartome\/","title":{"rendered":"The Isoaspartome"},"content":{"rendered":"

Deamidation of asparagine, Asn (and to a lesser extent glutamine, Gln) residues is the most common post-translation modification (PTM), occurring in many proteins as a non-enzymatic, spontaneous reaction. Deamidation plays an important role in aging, autoimmune disorders (e.g Celiac Disease), neurodegeneration (e.g. Alzheimer’s Disease), and other diseases; it can also influence the stability and potency of protein-based drugs. Under physiological conditions, Asn deamidation proceeds via a succinimide intermediate, which hydrolyzes to form a mixture of aspartate (Asp) and isoaspartate (isoAsp) residues.<\/p>\n

\"Asn<\/p>\n

Despite its ubiquity and importance in life science and in pharmaceutical industry, deamidation is underappreciated and understudied. This is in part due to the difficulty in differentiating the deamidation products, as Asp and isoAsp residues have the same mass and similar physiochemical properties. It was recently shown that electron capture dissociation (ECD) can cleave the C\u03b1<\/sub>-C\u03b2 <\/sub>bond in these residues resulting in diagnostic marker peaks in the MS\/MS spectra. For aspartic acid, it produces a M-60 peak, for isoaspartic acid, it produces c+57, z-57 peaks.<\/p>\n

\"Diagnostic<\/p>\n

To demonstrate this, a light chain peptide (46-61) from a patient with Primary Systemic Amyloidosis was subjected to ECD analysis, which generated isoAsp diagnostic z13<\/sub>-57 and Asp diagnostic M-60 ions, indicating the Asn49 residue has deamidated to form a mixture of Asp and isoAsp residues.<\/p>\n

\"\"<\/p>\n

Recent progresses of this work include the development of other electron activated dissociation methods, such as electron impact\/ionization dissociation (EID) and electron transfer dissociation (ETD) for isoAsp analysis, as well as for the related studies of glutamine deamidation (\u03b3-Glu identification). The potential of this method in the relative quantitation of isoAsp content, in the analysis of N-terminal isoAsp residues, and in analyzing HPLC-separated deamidation mixtures has been demonstrated. IsoAsp analysis employing top-down, middle-down, and MS3<\/sup> approaches is now possible. The ECD method has been successfully applied to monitor isoAsp formation in A-beta peptides, in proteins from patient samples, and in stressed protein-based drugs (e.g. Anthrax Vaccine). However,\u00a0ECD does not generate diagnostic ions for characterization of\u00a0\u03b2-amino acid residues other than isoAsp and \u03b2-phenylalanine due to the lack of radical stabilization.\u00a0We have recently showed that MALDI in-source decay (ISD) with a hydrogen-accepting matrix can\u00a0be broadly applied\u00a0for\u00a0\u03b2-peptide characterization by\u00a0producing site-specific a<\/em>-14 ions at\u00a0\u03b2-linkage sites.<\/span><\/p>\n

The following papers develop this work in detail:<\/p>\n