Open AccessLeveraging New Definitions of the LxVP SLiM To Discover Novel Calcineurin Regulators and Substrates
Author(s) -
Brooke L. Brauer,
Thomas M. Moon,
Sarah R. Sheftic,
Isha Nasa,
Rebecca Page,
Wolfgang Peti,
Arminja N. Kettenbach
Publication year - 2019
Publication title -
acs chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.899
H-Index - 111
eISSN - 1554-8937
pISSN - 1554-8929
DOI - 10.1021/acschembio.9b00606
Subject(s) - computational biology , phosphoprotein , proteome , binding site , complementarity (molecular biology) , calcineurin , sequence motif , plasma protein binding , interactome , biology , phosphatase , function (biology) , chemistry , biochemistry , microbiology and biotechnology , genetics , phosphorylation , gene , medicine , surgery , transplantation
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 S tructure B ased S hape C omplementarity (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.