BayesENproteomics: Bayesian elastic nets for quantification of proteoforms in complex samples

Abstract

Multivariate regression modelling provides a statistically powerful means of quantifying the effects of a given treatment while compensating for sources of variation and noise, such as variability between human donors and the behaviour of different peptides during mass spectrometry. However, methods to quantify endogenous post-translational modifications (PTMs) are typically reliant on summary statistical methods that fail to consider sources of variability such as changes in levels of the parent protein. Here, we compare three multivariate regression methods, including a novel Bayesian elastic net algorithm (BayesENproteomics) that enables assessment of relative protein abundances while also quantifying identified PTMs for each protein. We tested the ability of these methods to accurately quantify expression of proteins in a mixed-species benchmark experiment, and to quantify synthetic PTMs induced by stable isotope labelling. Finally, we extended our regression pipeline to calculate fold changes at the pathway level, providing a complement to commonly used enrichment analysis.Our results show that BayesENproteomics can quantify changes to protein levels across a broad dynamic range while also accurately quantifying PTM and pathway-level fold changes.

Laser capture microdissection coupled mass spectrometry (LCM-MS) for spatially resolved analysis of formalin-fixed and stained human lung tissues

“…Alignment decreases the number of missing values (a common problem in proteomics) at the risk of wrongly inferring presence of a peptide when it may genuinely be absent. Spectra from different tissue sections (human lung blood vessel or morphologically normal human lung alveoli) were analyzed either separately (with alignment between donors but not between sections; method that attempts to discern whether a given missing value is missing at random (MAR) or nonrandomly (MNR) and imputes from appropriate distributions (23). Reactome (24, 25) Pathway enrichment analysis was performed as described in (23), by fitting linear models for each pathway represented in the dataset, based on protein-level fold changes calculated as described above.…”