Crystal Structure of p38 Mitogen-activated Protein Kinase

Wilson, Keith; Fitzgibbon, Matthew J.; Caron, Paul L.; Griffith, Jeffrey K.; Chen, Wenyong; McCaffrey, Patricia G.; Chambers, Stephen P.; Su, Michael

doi:10.1074/jbc.271.44.27696

Cited by 239 publications

(195 citation statements)

References 30 publications

(34 reference statements)

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“…The classic mechanism of p38 activation is the phosphorylation of Thr 180 and Tyr 182 in the activation loop by the upstream dual specificity kinases MKK3 and MKK6. These phosphorylations in turn result in a rotation of the two lobes of p38, enabling Asp 168 in the C-terminal lobe and Lys 53 in the N-terminal lobe to reorient, thereby enabling their coordination in the binding and catalysis of ATP (21). Thus, in the absence of Thr 180 and Tyr 182 phosphorylation, it is unclear how the kinase can bind ATP and autophosphorylate.…”

Section: Discussionmentioning

confidence: 99%

A Chemical Genetic Approach Reveals That p38α MAPK Activation by Diphosphorylation Aggravates Myocardial Infarction and Is Prevented by the Direct Binding of SB203580

Kumphune

Bassi

Jacquet

et al. 2010

Journal of Biological Chemistry

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The use of nonselective pharmacological inhibitors has resulted in controversy regarding the mechanism and consequences of p38 activation during myocardial infarction. Classic p38 inhibitors such as SB203580 rely on a critical "gatekeeper" threonine residue for binding. We addressed these controversies by using mice in which the p38␣ alleles were targeted to cause substitution of the gatekeeper residue and resistance to inhibition. In homozygous drug-resistant compared with wildtype hearts, SB203580 failed to inhibit the activating phosphorylation of p38 or to reduce the infarction caused by myocardial ischemia. However, BIRB796, a p38 inhibitor not reliant on the gatekeeper for binding, similarly reduced p38-activating phosphorylation and infarction in both wild-type and knock-in mice, thereby excluding a nonspecific inhibitor-dependent phenotype resulting from the targeting strategy. Furthermore, the activation during myocardial ischemia involved phosphorylation of both the threonine and tyrosine residues in the activation loop of p38 despite the phosphorylation of the threonine alone being sufficient to create the epitope for dual phosphospecific antibody binding. Finally, SB203580 failed to reduce infarction in heterozygous drug-resistant hearts, suggesting that near complete inhibition of p38␣ kinase activity is necessary to elicit protection. These results indicate that, during myocardial ischemia, p38␣ (i) is the dominant-active p38 isoform, (ii) contributes to infarction, (iii) is responsible for the cardioprotective effect of SB203580, and (iv) is activated by a mechanism consistent with autodiphosphorylation despite this necessitating the phosphorylation of a tyrosine residue by an archetypal serine/threonine kinase.

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Section: Discussionmentioning

confidence: 99%

A Chemical Genetic Approach Reveals That p38α MAPK Activation by Diphosphorylation Aggravates Myocardial Infarction and Is Prevented by the Direct Binding of SB203580

Kumphune

Bassi

Jacquet

et al. 2010

Journal of Biological Chemistry

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“…The results obtained in the present study validate this approach. The activation loop differs significantly in sequence and structure between MAPKs and protein kinases in general (25,33,34) and is thus a handle of specificity. Furthermore, the 50-residue α-helical MAPK insertion and the extended C terminus (L16 segment) are unique to MAPKs and distinguish them from other members of the protein kinase family (24).…”

Section: Discussionmentioning

confidence: 99%

Structural and functional analysis of phosphorylation-specific binders of the kinase ERK from designed ankyrin repeat protein libraries

Kummer

Parizek

Rube

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

126

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We have selected designed ankyrin repeat proteins (DARPins) from a synthetic library by using ribosome display that selectively bind to the mitogen-activated protein kinase ERK2 (extracellular signal-regulated kinase 2) in either its nonphosphorylated (inactive) or doubly phosphorylated (active) form. They do not bind to other kinases tested. Crystal structures of complexes with two DARPins, each specific for one of the kinase forms, were obtained. The two DARPins bind to essentially the same region of the kinase, but recognize the conformational change within the activation loop and an adjacent area, which is the key structural difference that occurs upon activation. Whereas the rigid phosphorylated activation loop remains in the same form when bound by the DARPin, the more mobile unphosphorylated loop is pushed to a new position. The DARPins can be used to selectively precipitate the cognate form of the kinases from cell lysates. They can also specifically recognize the modification status of the kinase inside the cell. By fusing the kinase with Renilla luciferase and the DARPin to GFP, an energy transfer from luciferase to GFP can be observed in COS-7 cells upon intracellular complex formation. Phosphorylated ERK2 is seen to increase by incubation of the COS-7 cells with FBS and to decrease upon adding the ERK pathway inhibitor PD98509. Furthermore, the anti-ERK2 DARPin is seen to inhibit ERK phosphorylation as it blocks the target inside the cell. This strategy of creating activation-state-specific sensors and kinase-specific inhibitors may add to the repertoire to investigate intracellular signaling in real time.intrabodies | X-ray crystallography

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“…The structures of inactive and active (phosphorylated) p38␣ have been solved by X-ray crystallography. The phosphorylated TGY motif and the length of the activation loop were found to differ in ERK2 and JNK, which likely contributes to the substrate specificity of p38 (219,230).…”

Section: P38 Mapk Modulementioning

confidence: 99%

ERK and p38 MAPK-Activated Protein Kinases: a Family of Protein Kinases with Diverse Biological Functions

Roux

Blenis

2004

Microbiol Mol Biol Rev

2,129

1,861

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Conserved signaling pathways that activate the mitogen-activated protein kinases (MAPKs) are involved in relaying extracellular stimulations to intracellular responses. The MAPKs coordinately regulate cell proliferation, differentiation, motility, and survival, which are functions also known to be mediated by members of a growing family of MAPK-activated protein kinases (MKs; formerly known as MAPKAP kinases). The MKs are related serine/threonine kinases that respond to mitogenic and stress stimuli through proline-directed phosphorylation and activation of the kinase domain by extracellular signal-regulated kinases 1 and 2 and p38 MAPKs. There are currently 11 vertebrate MKs in five subfamilies based on primary sequence homology: the ribosomal S6 kinases, the mitogen- and stress-activated kinases, the MAPK-interacting kinases, MAPK-activated protein kinases 2 and 3, and MK5. In the last 5 years, several MK substrates have been identified, which has helped tremendously to identify the biological role of the members of this family. Together with data from the study of MK-knockout mice, the identities of the MK substrates indicate that they play important roles in diverse biological processes, including mRNA translation, cell proliferation and survival, and the nuclear genomic response to mitogens and cellular stresses. In this article, we review the existing data on the MKs and discuss their physiological functions based on recent discoveries

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Crystal Structure of p38 Mitogen-activated Protein Kinase

Cited by 239 publications

References 30 publications

A Chemical Genetic Approach Reveals That p38α MAPK Activation by Diphosphorylation Aggravates Myocardial Infarction and Is Prevented by the Direct Binding of SB203580

A Chemical Genetic Approach Reveals That p38α MAPK Activation by Diphosphorylation Aggravates Myocardial Infarction and Is Prevented by the Direct Binding of SB203580

Structural and functional analysis of phosphorylation-specific binders of the kinase ERK from designed ankyrin repeat protein libraries

ERK and p38 MAPK-Activated Protein Kinases: a Family of Protein Kinases with Diverse Biological Functions

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