Development of a comprehensive splice-site analysis using mass spectrometry-based proteomics

本研究旨在建立一种准确、全面地检测选择性剪接的剪接供体和受体部位的方法。该方法基于从头测序的方法，该方法直接从基于质谱学的蛋白质组数据中识别表达蛋白质的部分氨基酸序列。此外，为了提高剪接位点检测的准确性，新加入了利用多肽的理论物理化学性质来预测蛋白质组学中多肽检测的容易程度。虽然用蛋白质组学实验数据对所提出的方法进行了评估，但结果很差。然而，这导致了提高基于质谱学的蛋白质组数据的质量准确性的研究。提出了一种新的计算后验校正方法，并以裂解酵母蛋白质组数据为基础，证实了该方法提高了质量精度。

This study aimed to establish a method of accurately and comprehensively detecting the splice donor and acceptor sites of alternative splicing. The method is based on a de novo sequencing method, which directly identifies partial amino acid sequences of expressed proteins from the mass spectrometry-based proteome data. In addition, predicting the ease of peptide detection in proteomics using theoretical physicochemical properties of peptides is newly incorporated into the method in order to improve the accuracy of the splice site detection. Although the proposed method was evaluated with proteomics experimental data, it produced poor results. However, this led to the study of improving the mass accuracy of the mass spectrometry-based proteome data. A computational a posteriori calibration method was newly proposed, and based on the proteome data of fission yeast it was confirmed that the developed method increased mass accuracy.