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Curr Protoc Bioinformatics. 2015 Mar 9;49:8.19.1-16. doi: 10.1002/0471250953.bi0819s49.

Scoring Large-Scale Affinity Purification Mass Spectrometry Datasets with MiST.

Author information

  • 1Department of Cellular & Molecular Pharmacology, University of California, San Francisco, San Francisco, California; California Institute for Quantitative Biomedical Sciences, San Francisco, California.

Abstract

High-throughput Affinity Purification Mass Spectrometry (AP-MS) experiments can identify a large number of protein interactions, but only a fraction of these interactions are biologically relevant. Here, we describe a comprehensive computational strategy to process raw AP-MS data, perform quality controls, and prioritize biologically relevant bait-prey pairs in a set of replicated AP-MS experiments with Mass spectrometry interaction STatistics (MiST). The MiST score is a linear combination of prey quantity (abundance), abundance invariability across repeated experiments (reproducibility), and prey uniqueness relative to other baits (specificity). We describe how to run the full MiST analysis pipeline in an R environment and discuss a number of configurable options that allow the lay user to convert any large-scale AP-MS data into an interpretable, biologically relevant protein-protein interaction network.

KEYWORDS:

affinity purification mass spectrometry; interaction networks; protein interactions; proteomics; scoring algorithms

PMID:
25754993
PMCID:
PMC4378866
DOI:
10.1002/0471250953.bi0819s49
[PubMed - indexed for MEDLINE]
Free PMC Article
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