Large-scale proteoform analysis for unveiling the entire view

2023 Fiscal Year Final Research Report

• Project/Area Number: 21H02459
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 43050:Genome biology-related
• Research Institution: Kyoto University
• Principal Investigator: Ishihama Yasushi 京都大学, 薬学研究科, 教授 (30439244)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: プロテオフォーム / プロテオーム / スプライシング / 翻訳開始点 / プロテオリシス

Outline of Final Research Achievements

We developed the CHAMP method to isolate protein N- and C-terminal peptides from protein mixtures in two steps: digestion and isolation, respectively. The CHAMP method was applied to 11 non-small cell lung cancer cell lines and sample-selective proteoform profiles were measured. As a result, we successfully identified and quantified 3181 proteolysis products including unreported cases. In addition, 1122 peptides were identified as being derived from noncanonical proteins that are not included in the Swiss-Prot-derived proteins. Detailed analysis of the 1122 noncanonical protein-derived peptides led to the identification of many SNV-containing peptides, splicing isoforms, and novel translation start site candidates.

Free Research Field

プロテオミクス

Academic Significance and Societal Importance of the Research Achievements

近年タンパク質をnon-coding RNAが多数報告されてきたが、このなかには非典型タンパク質として翻訳されているものがあることがリボソームプロファイリングから予測されてきたが、今回の結果よりその全貌の一旦が明らかとなった。また、翻訳後にプロテオリシスをうけて生じたプロテオフォームについてもプロテオーム規模で明らかにすることができた。今後これらの制御機構がより詳細にわかり、疾患特異的なプロテオフォームが明らかになれば、新たな創薬標的となる可能性もある。