Home LiteratureArticle Details
PMID: 18239682 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Activation segment dimerization: a mechanism for kinase autophosphorylation of non-consensus sites.

The EMBO journal ·Vol. 27 ·No. 4 ·2008-02-20 ·Pages 704-14

Pike AC, Rellos P, Niesen FH, Turnbull A, Oliver AW, Parker SA, Turk BE, Pearl LH, Knapp S

Abstract

Protein kinase autophosphorylation of activation segment residues is a common regulatory mechanism in phosphorylation-dependent signalling cascades. However, the molecular mechanisms that guarantee specific and efficient phosphorylation of these sites have not been elucidated. Here, we report on three novel and diverse protein kinase structures that reveal an exchanged activation segment conformation. This dimeric arrangement results in an active kinase conformation in trans, with activation segment phosphorylation sites in close proximity to the active site of the interacting protomer. Analytical ultracentrifugation and chemical cross-linking confirmed the presence of dimers in solution. Consensus substrate sequences for each kinase showed that the identified activation segment autophosphorylation sites are non-consensus substrate sites. Based on the presented structural and functional data, a model for specific activation segment phosphorylation at non-consensus substrate sites is proposed that is likely to be common to other kinases from diverse subfamilies.

MeSH Terms
Dimerization Humans Models, Molecular Phosphorylation Protein Kinases/chemistry,metabolism Protein Structure, Tertiary
Chemicals
Protein Kinases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Pike Ashley C W
Structural Genomics Consortium, Botnar Research Centre, University of Oxford, Oxford, UK.
Rellos Peter
Niesen Frank H
Turnbull Andrew
Oliver Antony W
Parker Sirlester A
Turk Benjamin E
Pearl Laurence H
Knapp Stefan
References (46)
46 references, click to expand
  1. Regulation of zipper-interacting protein kinase activity in vitro and in vivo by multisite phosphorylation.
    J Biol Chem. 2005 Mar 11;280(10):9363-74 PMID: 15611134
  2. Structure and regulation of the human Nek2 centrosomal kinase.
    J Biol Chem. 2007 Mar 2;282(9):6833-42 PMID: 17197699
  3. A dimeric kinase assembly underlying autophosphorylation in the p21 activated kinases.
    J Mol Biol. 2006 Aug 11;361(2):312-26 PMID: 16837009
  4. Chemokine signaling and functional responses: the role of receptor dimerization and TK pathway activation.
    Annu Rev Immunol. 2001;19:397-421 PMID: 11244042
  5. Effects of phosphorylation of threonine 160 on cyclin-dependent kinase 2 structure and activity.
    J Biol Chem. 1999 Mar 26;274(13):8746-56 PMID: 10085115
  6. Activation-loop autophosphorylation is mediated by a novel transitional intermediate form of DYRKs.
    Cell. 2005 Jun 17;121(6):925-36 PMID: 15960979
  7. Phosphorylation of threonine 68 promotes oligomerization and autophosphorylation of the Chk2 protein kinase via the forkhead-associated domain.
    J Biol Chem. 2002 May 31;277(22):19389-95 PMID: 11901158
  8. Structural basis for recruitment of glycogen synthase kinase 3beta to the axin-APC scaffold complex.
    EMBO J. 2003 Feb 3;22(3):494-501 PMID: 12554650
  9. Chemical screening methods to identify ligands that promote protein stability, protein crystallization, and structure determination.
    Proc Natl Acad Sci U S A. 2006 Oct 24;103(43):15835-40 PMID: 17035505
  10. Death-associated protein kinase phosphorylates ZIP kinase, forming a unique kinase hierarchy to activate its cell death functions.
    Mol Cell Biol. 2004 Oct;24(19):8611-26 PMID: 15367680
  11. The Ste20-like kinase SLK is required for cell cycle progression through G2.
    J Biol Chem. 2005 Dec 23;280(51):42383-90 PMID: 16236704
  12. Discriminating between homodimeric and monomeric proteins in the crystalline state.
    Proteins. 2000 Oct 1;41(1):47-57 PMID: 10944393
  13. Oligomerization states of the association domain and the holoenyzme of Ca2+/CaM kinase II.
    FEBS J. 2006 Feb;273(4):682-94 PMID: 16441656
  14. Structures of human MAP kinase kinase 1 (MEK1) and MEK2 describe novel noncompetitive kinase inhibition.
    Nat Struct Mol Biol. 2004 Dec;11(12):1192-7 PMID: 15543157
  15. MOLPROBITY: structure validation and all-atom contact analysis for nucleic acids and their complexes.
    Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W615-9 PMID: 15215462
  16. ARP/wARP and automatic interpretation of protein electron density maps.
    Methods Enzymol. 2003;374:229-44 PMID: 14696376
  17. Structure of the autoinhibited kinase domain of CaMKII and SAXS analysis of the holoenzyme.
    Cell. 2005 Dec 2;123(5):849-60 PMID: 16325579
  18. Refinement of macromolecular structures by the maximum-likelihood method.
    Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55 PMID: 15299926
  19. MolProbity: all-atom contacts and structure validation for proteins and nucleic acids.
    Nucleic Acids Res. 2007 Jul;35(Web Server issue):W375-83 PMID: 17452350
  20. Macromolecular size-and-shape distributions by sedimentation velocity analytical ultracentrifugation.
    Biophys J. 2006 Jun 15;90(12):4651-61 PMID: 16565040
  21. The hCds1 (Chk2)-FHA domain is essential for a chain of phosphorylation events on hCds1 that is induced by ionizing radiation.
    J Biol Chem. 2001 Aug 10;276(32):30537-41 PMID: 11390408
  22. Stk10, a new member of the polo-like kinase kinase family highly expressed in hematopoietic tissue.
    J Biol Chem. 2003 May 16;278(20):18221-8 PMID: 12639966
  23. The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability.
    Nat Protoc. 2007;2(9):2212-21 PMID: 17853878
  24. A rapid method for determining protein kinase phosphorylation specificity.
    Nat Methods. 2004 Oct;1(1):27-9 PMID: 15782149
  25. Switching signals on or off by receptor dimerization.
    Cell. 1998 Aug 7;94(3):277-80 PMID: 9708728
  26. The importance of alignment accuracy for molecular replacement.
    Acta Crystallogr D Biol Crystallogr. 2004 Jul;60(Pt 7):1229-36 PMID: 15213384
  27. Crystal structure of the Src family tyrosine kinase Hck.
    Nature. 1997 Feb 13;385(6617):602-9 PMID: 9024658
  28. Structural basis for recruitment of the adaptor protein APS to the activated insulin receptor.
    Mol Cell. 2003 Dec;12(6):1379-89 PMID: 14690593
  29. DAPk protein family and cancer.
    Autophagy. 2006 Apr-Jun;2(2):74-9 PMID: 17139808
  30. LOSK (SLK) protein kinase activity is necessary for microtubule organization in the interphase cell centrosome.
    Dokl Biol Sci. 2005 Jul-Aug;403:317-9 PMID: 16358584
  31. THESEUS: maximum likelihood superpositioning and analysis of macromolecular structures.
    Bioinformatics. 2006 Sep 1;22(17):2171-2 PMID: 16777907
  32. Coot: model-building tools for molecular graphics.
    Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32 PMID: 15572765
  33. Mechanism of activation of ERK2 by dual phosphorylation.
    J Biol Chem. 2001 Jan 5;276(1):99-103 PMID: 11016942
  34. Regulation of protein kinases; controlling activity through activation segment conformation.
    Mol Cell. 2004 Sep 10;15(5):661-75 PMID: 15350212
  35. ZIP kinase, a novel serine/threonine kinase which mediates apoptosis.
    Mol Cell Biol. 1998 Mar;18(3):1642-51 PMID: 9488481
  36. A new vector for high-throughput, ligation-independent cloning encoding a tobacco etch virus protease cleavage site.
    Protein Expr Purif. 2002 Jun;25(1):8-15 PMID: 12071693
  37. Likelihood-enhanced fast rotation functions.
    Acta Crystallogr D Biol Crystallogr. 2004 Mar;60(Pt 3):432-8 PMID: 14993666
  38. Structure of mitogen-activated protein kinase-activated protein (MAPKAP) kinase 2 suggests a bifunctional switch that couples kinase activation with nuclear export.
    J Biol Chem. 2002 Oct 4;277(40):37401-5 PMID: 12171911
  39. Sequence analysis of the protein kinase gene family in human testicular germ-cell tumors of adolescents and adults.
    Genes Chromosomes Cancer. 2006 Jan;45(1):42-6 PMID: 16175573
  40. Crystal Structures of the p21-activated kinases PAK4, PAK5, and PAK6 reveal catalytic domain plasticity of active group II PAKs.
    Structure. 2007 Feb;15(2):201-13 PMID: 17292838
  41. Trans-activation of the DNA-damage signalling protein kinase Chk2 by T-loop exchange.
    EMBO J. 2006 Jul 12;25(13):3179-90 PMID: 16794575
  42. Structural basis for the autoinhibition of calcium/calmodulin-dependent protein kinase I.
    Cell. 1996 Mar 22;84(6):875-87 PMID: 8601311
  43. A chaperone-dependent GSK3beta transitional intermediate mediates activation-loop autophosphorylation.
    Mol Cell. 2006 Nov 17;24(4):627-33 PMID: 17188038
  44. Activation segment exchange: a common mechanism of kinase autophosphorylation?
    Trends Biochem Sci. 2007 Aug;32(8):351-6 PMID: 17627826
  45. Two-stage mechanism for activation of the DNA replication checkpoint kinase Cds1 in fission yeast.
    Genes Dev. 2006 Apr 15;20(8):990-1003 PMID: 16618806
  46. Pyridone 6, a pan-Janus-activated kinase inhibitor, induces growth inhibition of multiple myeloma cells.
    Cancer Res. 2006 Oct 1;66(19):9714-21 PMID: 17018630
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
DOI
10.1038/emboj.2008.8
Published
2008-02-20
Epub
2008-00-31
Pages
704-14
Language
English
Region
England
NLM ID
8208664
PMCID
PMC2239268
Subset
IM
Grants
Wellcome Trust · United Kingdom
NIGMS NIH HHS · R01 GM079498 · United States
NIGMS NIH HHS · GM079498 · United States
Databases
PDB
2J51, 2J7T, 2J90, 2JFL, 2JFM, 2UV2
External Links
PubMed source DOI: 10.1038/emboj.2008.8 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2