演題概要

若手研究者セッション

第2日　5月16日（火）　09:45～10:00　C会場（会議室1009）

2C-O-0945（2P-18）　　PDF

ピペットチップ型カラムを用いたワンステップタンパク質末端ペプチド濃縮法

(¹京大院薬・ ²理研IMS・ ³医薬基盤健栄研)
^o森川和哉¹・ 西田紘士¹・ 今見考志^1,2・ 石濱泰^1,3

Comprehensive profiling of protein termini is important for understanding the function of proteoforms. In this study, we developed a new protocol for the CHAMP-N method, a one-step enrichment of protein N-terminal peptides by strong cation exchange chromatography (SCX) for protein digested peptides by LysargiNase. In this new protocol, a disposable pipette-tip SCX microcolumn is used instead of a conventional HPLC column, allowing for simplified, parallel processing and high-throughput analysis of protein N-terminal peptides. Furthermore, by optimizing the elution conditions, more protein N-terminal peptides were successfully identified than in the conventional HPLC-based method. In this presentation, we will also report the results of comprehensive analysis of protein N- and C-terminal peptides using our recently developed C-terminal peptide enrichment method, CHAMP-C, combined with the CHAMP-N method.