Oral Sessions
- Day 1, May 11（Mon.） 15:50-16:10 Room A (Main Hall)
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1A-O1-1550 PDF
The use of barcode-peptide tags toward proteomic analysis with high sensitivity
In mass spectrometry (MS)-based proteomic analysis, an advance of analytical platform has achieved a variety of proteome-wide analyses. However, it is still difficult to analyze the whole proteome even in the budding yeast, a simple model organism. This would be not only because of very low expression levels of some missing proteins, but also due to a large variation of individual proteins sequences. Actually, detection sensitivities of individual tryptic peptides are quite different with thousands or more depending on each sequence and chemical property. Therefore, some proteins have few peptides suited to be detected by MS analysis. To overcome this problem, we conducted a strategy in which budding yeast, used a model organism, is genetically modified to be successful for MS-analysis of some proteins that are intrinsically difficult to detect. In brief, barcode peptide tags each having unique sequence and easy to detect with high sensitivity are introduced to target proteins to be analyzed. We have screened the sequences for peptide tags, then created a yeast strain where unique peptide tag are integrated into multiple proteins by the use of CRISPR/Cas9 system. Here we will present a proof-of-concept of this strategy and also perspectives of its applicability.