A Bioconductor workflow for processing, evaluating, and interpreting expression proteomics data.
Charlotte HutchingsCharlotte S DawsonThomas KruegerKathryn S LilleyLisa M BreckelsPublished in: F1000Research (2023)
Background: Expression proteomics involves the global evaluation of protein abundances within a system. In turn, differential expression analysis can be used to investigate changes in protein abundance upon perturbation to such a system. Methods: Here, we provide a workflow for the processing, analysis and interpretation of quantitative mass spectrometry-based expression proteomics data. This workflow utilizes open-source R software packages from the Bioconductor project and guides users end-to-end and step-by-step through every stage of the analyses. As a use-case we generated expression proteomics data from HEK293 cells with and without a treatment. Of note, the experiment included cellular proteins labelled using tandem mass tag (TMT) technology and secreted proteins quantified using label-free quantitation (LFQ). Results: The workflow explains the software infrastructure before focusing on data import, pre-processing and quality control. This is done individually for TMT and LFQ datasets. The application of statistical differential expression analysis is demonstrated, followed by interpretation via gene ontology enrichment analysis. Conclusions: A comprehensive workflow for the processing, analysis and interpretation of expression proteomics is presented. The workflow is a valuable resource for the proteomics community and specifically beginners who are at least familiar with R who wish to understand and make data-driven decisions with regards to their analyses.
Keyphrases
- mass spectrometry
- label free
- electronic health record
- poor prognosis
- binding protein
- liquid chromatography
- data analysis
- long non coding rna
- healthcare
- gas chromatography
- big data
- capillary electrophoresis
- small molecule
- cell proliferation
- ms ms
- genome wide
- gene expression
- microbial community
- dna methylation
- tandem mass spectrometry
- liquid chromatography tandem mass spectrometry
- single cell
- amino acid
- cell cycle arrest
- pi k akt
- solid phase extraction