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Leveraging New Definitions of the LxVP SLiM To Discover Novel Calcineurin Regulators and Substrates
journal contributionposted on 2019-11-01, 18:39 authored by Brooke L. Brauer, Thomas M. Moon, Sarah R. Sheftic, Isha Nasa, Rebecca Page, Wolfgang Peti, Arminja N. Kettenbach
The Phosphoprotein Phosphatase Calcineurin (CN, PP2B, PP3) recognizes and binds to two short linear motifs (SLiMs), PxIxIT and LxVP, in its regulators and substrates. These interactions enable CN function in many key biological processes. The identification of SLiMs is difficult because of their short, degenerate sequence and often low binding affinity. Here we combine Structure Based Shape Complementarity (SBSC) analysis and proteome-wide affinity purification-mass spectrometry to identify PxIxIT and LxVP containing CN interactors to expand and thereby redefine the LxVP motif. We find that the new πφ-LxVx primary sequence defines an ensemble of binding competent confirmations and thus the binding on-rate, making it difficult to predict the LxVP binding strength from its sequence. Our analysis confirms existing and, more importantly, identifies novel CN interactors, substrates, and thus biological functions of CN.
sequenceCN interactorsCN functionS tructure B ased S hape C omplementarityPPLxVP binding strengthDiscover Novel Calcineurin Regulatorsπφ- LxVxPhosphoprotein Phosphatase CalcineurinLeveraging New Definitions2Bproteome-wide affinity purification-mass spectrometrynovel CN interactorsSBSCsubstratePxIxITanalysisbinding affinityLxVP SLiMLxVP motifbinding on-rate