Analysis of Arginylated Peptides by Subtractive Edman Degradation

  • Anna S. Kashina
  • John R. YatesIIIEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1337)


During the early studies of N-terminal arginylation, Edman degradation was widely used to identify N-terminally added Arg on protein substrates. This old method is reliable but highly depends on the purity and abundance of samples and can become misleading unless a highly purified, highly arginylated protein can be obtained. Here we report a mass spectrometry-based method that utilizes Edman degradation chemistry to identify arginylation in more complex and less abundant protein samples. This method can also apply to the analysis of other posttranslational modifications.

Key words

Edman degradation Mass spectrometry Arginylation 


  1. 1.
    Bradley CV, Williams DH, Hanley MR (1982) Peptide sequencing using the combination of edman degradation, carboxypeptidase digestion and fast atom bombardment mass spectrometry. Biochem Biophys Res Commun 104(4):1223–1230CrossRefPubMedGoogle Scholar
  2. 2.
    Hunt DF, Yates JR III, Shabanowitz J, Winston S, Hauer CR (1986) Protein sequencing by tandem mass spectrometry. Proc Natl Acad Sci U S A 83(17):6233–6237PubMedCentralCrossRefPubMedGoogle Scholar
  3. 3.
    Wang J, Han X, Wong CC, Cheng H, Aslanian A, Xu T, Leavis P, Roder H, Hedstrom L, Yates JR III, Kashina A (2014) Arginyltransferase ATE1 catalyzes midchain arginylation of proteins at side chain carboxylates in vivo. Chem Biol 21(3):331–337PubMedCentralCrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Animal Biology, School of Veterinary MedicineUniversity of PennsylvaniaPhiladelphiaUSA
  2. 2.Department of Chemical PhysiologyThe Scripps Research InstituteLa JollaUSA

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