Published in

American Chemical Society, Journal of Proteome Research, 3(11), p. 1462-1474, 2012

DOI: 10.1021/pr2011632

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Assignment of Protein Interactions from Affinity Purification/Mass Spectrometry Data

Journal article published in 2012 by Mercedes Pardo ORCID, Jyoti S. Choudhary ORCID
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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Abstract

The combination of affinity purification with mass spectrometry analysis has become the method of choice for protein complex characterization. With the improved performance of mass spectrometry technology, the sensitivity of the analyses is increasing, probing deeper into molecular interactions and yielding longer lists of proteins. These identify not only core complex subunits but also the more inaccessible proteins that interact weakly or transiently. Alongside them, contaminant proteins, which are often abundant proteins in the cell, tend to be recovered in affinity experiments because they bind nonspecifically and with low affinity to matrix, tag, and/or antibody. The challenge now lies in discriminating nonspecific binders from true interactors, particularly at the low level and in a larger scale. This review aims to summarize the variety of methods that have been used to distinguish contaminants from specific interactions in the past few years, ranging from manual elimination using heuristic rules to more sophisticated probabilistic scoring approaches. We aim to give awareness on the processing that takes place before an interaction list is reported and on the different types of list curation approaches suited to the different experiments.