title: Regulatory roles of conserved phosphorylation sites in the activation T-loop of the MAP kinase ERK1.
abstract: The catalytic domains of most eukaryotic protein kinases are highly conserved in their primary structures. Their phosphorylation within the well-known activation T-loop, a variable region between protein kinase catalytic subdomains VII and VIII, is a common mechanism for stimulation of their phosphotransferase activities. Extracellular signal-regulated kinase 1 (ERK1), a member of the extensively studied mitogen-activated protein kinase (MAPK) family, serves as a paradigm for regulation of protein kinases in signaling modules. In addition to the well-documented T202 and Y204 stimulatory phosphorylation sites in the activation T-loop of ERK1 and its closest relative, ERK2, three additional flanking phosphosites have been confirmed (T198, T207, and Y210 from ERK1) by high-throughput mass spectrometry. In vitro kinase assays revealed the functional importance of T207 and Y210, but not T198, in negatively regulating ERK1 catalytic activity. The Y210 site could be important for proper conformational arrangement of the active site, and a Y210F mutant could not be recognized by MEK1 for phosphorylation of T202 and Y204 in vitro. Autophosphorylation of T207 reduces the catalytic activity and stability of activated ERK1. We propose that after the activation of ERK1 by MEK1, subsequent slower phosphorylation of the flanking sites results in inhibition of the kinase. Because the T207 and Y210 phosphosites of ERK1 are highly conserved within the eukaryotic protein kinase family, hyperphosphorylation within the kinase activation T-loop may serve as a general mechanism for protein kinase down-regulation after initial activation by their upstream kinases.
authors.affiliation: Division of Neurology, Department of Medicine, University of British Columbia, Vancouver, BC V6T 2B5, Canada.
authors.forename: Shenshen
authors.initials: S
authors.lastname: Lai
authors.affiliation: Division of Neurology, Department of Medicine, University of British Columbia, Vancouver, BC V6T 2B5, Canada Kinexus Bioinformatics Corporation, Vancouver, BC V6P 6T3, Canada spelech@kinexus.ca.
authors.forename: Steven
authors.initials: S
authors.lastname: Pelech
mesh_terms: D000818:Animals; D020134:Catalytic Domain; D002460:Cell Line; D015536:Down-Regulation; D006801:Humans; D048369:MAP Kinase Kinase 1 / Q000378:metabolism; D020935:MAP Kinase Signaling System; D048052:Mitogen-Activated Protein Kinase 3 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation
chemical_list: D048052:Mitogen-Activated Protein Kinase 3; D048369:MAP Kinase Kinase 1
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