1. Deep-Learning-Derived Evaluation Metrics Enable Effective Benchmarking of Computational Tools for Phosphopeptide Identification
- Author
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Vladislav A. Petyuk, Bing Zhang, Felipe da Veiga Leprevost, Wen Jiang, Alexey I. Nesvizhskii, Wen-Feng Zeng, Nathan Edwards, Tao Liu, Kai Li, Jamie Moon, and Bo Wen
- Subjects
Phosphopeptides ,Proteomics ,RT, retention time ,False discovery rate ,MS/MS, tandem mass spectrometry ,SPC, Spearman’s correlation coefficient ,FDR, false discovery rate ,Computer science ,NL, neutral loss ,CDAP, CPTAC common data analysis pipeline ,Benchmark ,PCC, Pearson’s correlation coefficient ,Tandem mass tag ,computer.software_genre ,NCE, normalized collision energy ,Cell Line ,Set (abstract data type) ,Mice ,Deep Learning ,COS, cosine similarity ,Animals ,Humans ,Phosphorylation ,phosphopeptide identification ,SA, spectral angle ,LC, liquid chromatography ,Research ,Cosine similarity ,CPTAC, Clinical Proteomic Tumor Analysis Consortium ,Phosphoproteomics ,phosphoproteomics ,TMT, tandem mass tag ,General Medicine ,Benchmarking ,PSM, peptide-spectrum match ,ICPC, International Cancer Proteogenome Consortium ,AUROC, area under the receiver operating characteristics ,Identification (information) ,UCEC, uterine corpus endometrial carcinoma ,ComputingMethodologies_PATTERNRECOGNITION ,MAE, median absolute error ,KDT, Kendal rank correlation coefficient ,Benchmark (computing) ,PTM, posttranslational modification ,Data mining ,computer - Abstract
Tandem mass spectrometry (MS/MS)-based phosphoproteomics is a powerful technology for global phosphorylation analysis. However, applying four computational pipelines to a typical mass spectrometry (MS)-based phosphoproteomic dataset from a human cancer study, we observed a large discrepancy among the reported phosphopeptide identification and phosphosite localization results, underscoring a critical need for benchmarking. While efforts have been made to compare performance of computational pipelines using data from synthetic phosphopeptides, evaluations involving real application data have been largely limited to comparing the numbers of phosphopeptide identifications due to the lack of appropriate evaluation metrics. We investigated three deep-learning-derived features as potential evaluation metrics: phosphosite probability, Delta RT, and spectral similarity. Predicted phosphosite probability is computed by MusiteDeep, which provides high accuracy as previously reported; Delta RT is defined as the absolute retention time (RT) difference between RTs observed and predicted by AutoRT; and spectral similarity is defined as the Pearson’s correlation coefficient between spectra observed and predicted by pDeep2. Using a synthetic peptide dataset, we found that both Delta RT and spectral similarity provided excellent discrimination between correct and incorrect peptide-spectrum matches (PSMs) both when incorrect PSMs involved wrong peptide sequences and even when incorrect PSMs were caused by only incorrect phosphosite localization. Based on these results, we used all the three deep-learning-derived features as evaluation metrics to compare different computational pipelines on diverse set of phosphoproteomic datasets and showed their utility in benchmarking performance of the pipelines. The benchmark metrics demonstrated in this study will enable users to select computational pipelines and parameters for routine analysis of phosphoproteomics data and will offer guidance for developers to improve computational methods., Graphical Abstract, Highlights • Computational method selection substantially affects phosphopeptide identification. • Deep-learning-derived metrics effectively discriminate correct and incorrect PSMs. • Novel metrics enable computational method comparison on real application data., In Brief Tandem mass spectrometry (MS/MS)-based phosphoproteomics is a powerful technology for global phosphorylation analysis. However, applying different computational pipelines to the same dataset may produce substantially different phosphopeptide identification results, underscoring a critical need for benchmarking. We present three deep-learning-derived benchmark metrics. The benchmark metrics demonstrated in this study will enable users to select computational pipelines and parameters for routine analysis of phosphoproteomics data and will offer guidance for developers to improve computational methods.
- Published
- 2021