title: Structural Studies of ERK2 Protein Complexes.
abstract: ERK1 and ERK2 (ERK1/2) are the primary effector kinases of the RAS-RAF-MEK-ERK signaling pathway. A variety of substrates and regulatory partners associate with ERK1/2 through distinct D-peptide- and DEF-docking sites on their kinase domains. While understanding of D-peptides that bind to ERK1/2 has become increasingly clear over the last decade, only more recently have structures of proteins interacting with other binding sites on ERK1/2 become available. PEA-15 is a 130-residue ERK1/2 regulator that engages both the D-peptide- and DEF-docking sites of ERK kinases, and directly sequesters the ERK2 activation loop in various different phosphorylation states. Here we describe the methods used to derive crystallization-grade complexes of ERK2-PEA-15, which may also be adapted for other regulators that associate with the activation loop of ERK1/2.
authors.affiliation: Biochemistry Department, Otago School of Medical Sciences, University of Otago, 56, 710 Cumberland St., Dunedin, 9054, New Zealand.
authors.forename: Johannes F
authors.initials: JF
authors.lastname: Weijman
authors.affiliation: Cell Death and Survival Networks Program, Sanford Burnham Prebys Medical Discovery Institute, 10901 North Torrey Pines Road, La Jolla, CA, 92037, USA.
authors.forename: Stefan J
authors.initials: SJ
authors.lastname: Riedl
authors.affiliation: Biochemistry Department, Otago School of Medical Sciences, University of Otago, 56, 710 Cumberland St., Dunedin, 9054, New Zealand. peter.mace@otago.ac.nz.
authors.forename: Peter D
authors.initials: PD
authors.lastname: Mace
mesh_terms: D051017:Apoptosis Regulatory Proteins; D001665:Binding Sites; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins; D025842:SUMO-1 Protein / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D051017:Apoptosis Regulatory Proteins; D047908:Intracellular Signaling Peptides and Proteins; D046912:Multiprotein Complexes; C092039:PEA15 protein, human; D010750:Phosphoproteins; D011994:Recombinant Proteins; D025842:SUMO-1 Protein; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1
keywords: Activation loop; Crystallization; D-Peptide; DEF-docking; DEJL; ERK1; ERK2; MAPK; PEA-15