Program

Poster Presentations

Day 3, May 17（Thu.）　　Poster

3P-03　　PDF

Investigation for a novel C-terminal sequencing method

(RIKEN CSRS)
^oTakehiro Suzuki, Naoshi Dohmae

It is well known that the C-terminus of a protein undergoes processing containing protease hydrolysis and various post-translational modifications. Although the C-terminal analysis has been studied for a long time, the definitive method has not been developed yet despite its biological importance. To sequence the C-terminus of a protein or peptide, a method of sequentially hydrolyzing the C-terminus is used. The C-terminal amino acid sequence is determined by measuring the obtained peptide mixture or amino acid. However, the existing methods have not come to be widely used due to the specificity of enzymes, the complication of chemical reactions and low sensitivity.
To solve the above problems, we considered the combination of a novel protein hydrolyzation and mass spectrometry. We applied the protein degradation reaction caused in the presence of copper ions and ascorbic acid to C-terminal sequencing. We were able to find ladder peaks of the C-terminal region with the oxidized insulin B chain, Myoglobin and Ubiquitin. In this research, we found the metal ion and ascorbic acid caused the degradation of the C-terminal region of the protein, and the mass spectrometric measurement of the resultant peptides enabled us to determine C-terminal sequence.