Abstract
To study the mechanism of protein carboxyl methyltransferase-driven repair of age-damaged sites in polypeptides, a modell-isoaspartyl peptide,l-isotetragastrin, was enzymatically repaired to normall-tetragastrin in the presence of18O-enriched water. By this design, the enrichment of18O atoms in the peptide would reflect the number of passages through a hydrolyzable succinimide intermediate during formation of the repaired product. Mass determinations by FAB mass spectrometry revealed repaired peptide with two18O atoms incorporated, demonstrating that more than a single cycle of methylation and demethylation is necessary to ensure stoichiometric repair.
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Abbreviations
- HPLC:
-
high-pressure liquid chromatography
- FAB:
-
fast atom bombardment
- TFA:
-
trifluoroacetic acid
- PCM:
-
proteind-aspartyl/L-isoaspartyl carboxyl methyltransfer-ase
- l-Normal:
-
[l-Asp3]tetragastrin
- l-Iso:
-
[L-isoAsp3]tetragastrin
- d-Normal:
-
[d-Asp3]tetragastrin
- d-Iso:
-
[d-isoAsp3]tetragastrin
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Lindquist, J.A., McFadden, P.N. Incorporation of two18O atoms into a peptide during isoaspartyl repair reveals repeated passage through a succinimide intermediate. J Protein Chem 13, 553–560 (1994). https://doi.org/10.1007/BF01901537
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DOI: https://doi.org/10.1007/BF01901537