title: Interconversion of the peptide isoforms of aspartate: stability of isoaspartates.
abstract: A common modification of human long-lived proteins is spontaneous isomerisation of aspartate residues, and its biological importance can be inferred from the ubiquitous presence of protein isoaspartate methyl transferase (PIMT), that repairs this damage. Cyclisation of L-Asp residues yields four isomers: L-Asp, L-isoAsp, D-Asp and D-isoAsp, however little is known about their rate of formation or interconversion. This is important because PIMT is inactive towards D-isoAsp. Peptides containing the four Asp isoforms corresponding to a susceptible site (Asp 151) in the chaperone, αA-crystallin, were examined for their interconversion at pH 7. D-Asp formed from L-Asp readily, whereas L-isoAsp was not detected until significantly later. D-isoAsp formed very slowly, with just 1% present after 8 days at 60°C. These findings can be used to rationalise the substrate specificity of PIMT. In addition, both the D-isoAsp and L-isoAsp peptides were found to be remarkably stable, showing little conversion to other isomers, even after weeks of incubation. Therefore L-isoAsp and D-isoAsp appear to represent "terminal" stages of L-Asp modification. If PIMT is present, L-isoAsp may be reverted to L-Asp, however there appears to be no prospect of reversing D-isoAsp formation in aged proteins. Interestingly, Asp 151 in recombinant αA crystallin isomerised more rapidly than in the L-Asp peptide.
authors: Save Sight Institute, Sydney Eye Hospital, University of Sydney, 8 Macquarie Street, Sydney, New South Wales 2000, Australia.
authors: Michelle Y S
authors: MY
authors: Hooi
authors: Mark J
authors: MJ
authors: Raftery
authors: Roger J W
authors: RJ
authors: Truscott
mesh_terms: D001224:Aspartic Acid / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D026581:Isoaspartic Acid / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D026601:Protein D-Aspartate-L-Isoaspartate Methyltransferase / Q000378:metabolism*; D020033:Protein Isoforms; D017433:Protein Structure, Secondary; D013379:Substrate Specificity; D013997:Time Factors; D014357:Trypsin / Q000737:chemistry; D038202:alpha-Crystallin A Chain / Q000737:chemistry
chemical_list: D026581:Isoaspartic Acid; D010455:Peptides; D020033:Protein Isoforms; D038202:alpha-Crystallin A Chain; D001224:Aspartic Acid; D026601:Protein D-Aspartate-L-Isoaspartate Methyltransferase; D014357:Trypsin