Analysis of Limited Proteolysis-Coupled Mass Spectrometry Data


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Date

2025-04

Publication Type

Journal Article

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yes

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Abstract

Limited proteolysis combined with mass spectrometry (LiP-MS) facilitates probing structural changes on a proteome-wide scale. This method leverages differences in the proteinase K accessibility of native protein structures to concurrently assess structural alterations for thousands of proteins in situ. Distinguishing different contributions to the LiP-MS signal, such as changes in protein abundance or chemical modifications, from structural protein alterations remains challenging. Here, we present the first comprehensive computational pipeline to infer structural alterations for LiP-MS data using a two-step approach. 1) We remove unwanted variations from the LiP signal that are not caused by protein structural effects and 2) infer the effects of variables of interest on the remaining signal. Using LiPMS data from three species, we demonstrate that this approach outperforms previously employed approaches. Our framework provides a uniquely powerful approach for deconvolving LiP-MS signals and separating protein structural changes from changes in protein abundance, posttranslational modifications, and alternative splicing. Our approach may also be applied to analyze other types of peptide-centric structural proteomics data, such as FPOP or molecular painting data.

Publication status

published

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Book title

Volume

24 (4)

Pages / Article No.

100934

Publisher

Elsevier

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Edition / version

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Software

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Organisational unit

03927 - Picotti, Paola / Picotti, Paola check_circle

Notes

Funding

866004 - Three-dimensional dynamic views of proteomes as a novel readout for physiolgical and pathological alterations (EC)

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