Mapping ERK2-MKP3 binding interfaces by hydrogen/deuterium exchange mass spectrometry

Bo Zhou, Jialin Zhang, Sijiu Liu, Sharanya Reddy, Fang Wang, Zhong Yin Zhang

Research output: Contribution to journalArticle

42 Scopus citations

Abstract

ERK2, a prototypic member of the MAPK family, plays a central role in regulating cell growth and differentiation. MKP3, an ERK2-specific phosphatase, terminates ERK2 signaling. To understand the molecular basis of ERK2 recognition by MKP3, we carried out hydrogen/deuterium exchange mass spectrometry experiments to map the interaction surfaces between the two proteins. The results show that the exquisite specificity of MKP3 for ERK2 is governed by two distinctive protein-protein interactions. To increase the "effective concentration" of the interacting molecules, the kinase interaction motif in MKP3 (64RRLQKGNLPVR74) and an MKP3-specific segment (101NSSDWNE107) bind the common docking site in ERK2 defined by residues in L16, L5, β7- β8, and αd-L8e, located opposite the kinase active site. In addition to this "tethering" effect, additional interactions between the 364FTAP367 sequence in MKP3 and the ERK2 substrate-binding site, formed by residues in the activation lip and the P+1 site (β9f loop), L13fg loop), and the MAPK insert (L 141L142L14), are essential for allosteric activation of MKP3 and formation of a productive complex whereby the MKP3 catalytic site is correctly juxtaposed to carry out the dephosphorylation of phospho-Thr183/phospho-Tyr185 in ERK2. This bipartite protein-protein interaction model may be applicable to the recognition of other MAPKs by their cognate regulators and substrates.

Original languageEnglish (US)
Pages (from-to)38834-38844
Number of pages11
JournalJournal of Biological Chemistry
Volume281
Issue number50
DOIs
StatePublished - Dec 15 2006

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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