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Structural Studies of ERK2 Protein Complexes

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Book cover ERK Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1487))

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.

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Acknowledgements

This work was supported by NIH grants R01AA017238, 1R01CA160457 (to S.J.R.), and a DOD-BCRP Fellowship BC100466 to P.D.M. P.D.M. and J.F.W. are currently supported by a Rutherford Discovery Fellowship from the New Zealand government administered by the Royal Society of New Zealand (to P.D.M.). The original pET-LIC vector used in this work was kindly gifted by the Netherlands Cancer Institute (NKI) Protein Facility with funding from grant no. 175.010.2007.012.

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Authors and Affiliations

  1. Biochemistry Department, Otago School of Medical Sciences, University of Otago, 56, 710 Cumberland St., Dunedin, 9054, New Zealand

    Johannes F. Weijman & Peter D. Mace

  2. Cell Death and Survival Networks Program, Sanford Burnham Prebys Medical Discovery Institute, 10901 North Torrey Pines Road, La Jolla, CA, 92037, USA

    Stefan J. Riedl

Authors
  1. Johannes F. Weijman
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  2. Stefan J. Riedl
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  3. Peter D. Mace
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Corresponding author

Correspondence to Peter D. Mace .

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Editors and Affiliations

  1. Institut de Biologia Molecular de Barcelona-CSIC, Parc Científic de Barcelona, Barcelona, Spain

    Gerardo Jimenez

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Weijman, J.F., Riedl, S.J., Mace, P.D. (2017). Structural Studies of ERK2 Protein Complexes. In: Jimenez, G. (eds) ERK Signaling. Methods in Molecular Biology, vol 1487. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6424-6_4

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  • DOI: https://doi.org/10.1007/978-1-4939-6424-6_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6422-2

  • Online ISBN: 978-1-4939-6424-6

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