Applying Arginylation for Bottom-Up Proteomics

  • H. Alexander EbhardtEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1337)


Arginylation is an enzymatic reaction in which arginyl-tRNA protein transferase 1 (ATE1, EC conjugates a single arginyl moiety from aminoacylated tRNAArg onto a target polypeptide. We established arginylation for in vitro labeling of peptides with N-terminal acidic amino acids. Consistent with prior knowledge, arginylated peptides flanked by basic amino acids result in rich redundant MS/MS fragment spectra using various precursor fragmentation modes. Arginylation carried out by ATE1 is a fast method for labeling peptides. Sequence-specific proteolytic digest of proteins is best carried out using a double digest of proteins by Lys-C and Asp-N to generate peptides with a basic amino acid on the C-terminus and an acidic amino acid on the N-terminus. Under these conditions, arginylation is specific for N-terminal acidic amino acids and results in a near 2× sequence coverage in the MS/MS spectrum are achieved.

Key words

Selected reaction monitoring (SRM) Multiple reaction monitoring (MRM) Mass spectrometry Arginylation 



This work was supported by a FP7 Marie Curie International Incoming Fellowship to HAE.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Institute of Molecular Systems BiologyEidgenössische Technische Hochschule (ETH) ZürichZürichSwitzerland

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