The structural basis for the specificity of pyridinylimidazole inhibitors of p38 MAP kinase

Wilson, Keith; McCaffrey, Patricia G.; Hsiao, Kathy; Pazhanisamy, S.; Galullo, Vincent; Bemis, Guy W.; Fitzgibbon, Matthew J.; Caron, Paul L.; Murcko, Mark A.; Su, Michael

doi:10.1016/s1074-5521(97)90194-0

Cited by 274 publications

(242 citation statements)

References 31 publications

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“…As shown in Figure 2A, the MEK/ERK inhibitor PD98059 [15] diminished the anti-exocytotic effect of imetit by ~30% at 3 μM and abolished it at 10 μM. Similarly, the p38 inhibitor SB202190 [16] diminished the effect of imetit by ~60% at 30 nM and abolished it at 100 nM ( Fig. 2B), while the JNK inhibitor SP600125 [17] diminished the effect of imetit by ~30% at 150 nM and abolished it at 200 nM ( Fig.…”

Section: Mapk Activation Plays a Role In The H 3 R-induced Inhibitionmentioning

confidence: 69%

Histamine H3-receptor signaling in cardiac sympathetic nerves: Identification of a novel MAPK-PLA2-COX-PGE2-EP3R pathway

Levi

Seyedi

Schäefer

et al. 2007

Biochemical Pharmacology

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We tested the hypothesis that the histamine H 3 -receptor (H 3 R)-mediated attenuation of norepinephrine (NE) exocytosis from cardiac sympathetic nerves results not only from a Gα imediated inhibition of the adenylyl cyclase-cAMP-PKA pathway, but also from a Gβγ i -mediated activation of the MAPK-PLA 2 cascade, culminating in formation of an arachidonate metabolite with anti-exocytotic characteristics (e.g., PGE 2 ). We report in Langendorff-perfused guinea-pig hearts and isolated sympathetic nerve endings (cardiac synaptosomes), H 3 R-mediated attenuation of K + -induced NE exocytosis was prevented by MAPK and PLA 2 inhibitors, and by cyclooxygenase and EP 3 -receptor (EP 3 R) antagonists. Moreover, H 3 R activation resulted in MAPK phosphorylation in H 3 R-transfected SH-SY5Y neuroblastoma cells, and in PLA 2 activation and PGE 2 production in cardiac synaptosomes; H 3 R-induced MAPK phosphorylation was prevented by an anti-βγ peptide. Synergism between H 3 R and EP 3 R agonists (i.e., imetit and sulprostone, respectively) suggested PGE 2 may be a downstream effector of the anti-exocytotic effect of H 3 R activation. Furthermore, the anti-exocytotic effect of imetit and sulprostone was potentiated by the N-type Ca 2+ -channel antagonist ω-conotoxin GVIA, and prevented by an anti-Gβγ peptide. Our findings suggest an EP 3 R Gβγ i -induced decrease in Ca 2+ influx through N-type Ca 2+ -channels is involved in PGE 2 / EP 3 R-mediated attenuation of NE exocytosis elicited by H 3 R activation. Conceivably, activation of the Gβγ i subunit of H 3 R and EP 3 R may also inhibit Ca 2+ entry directly, independent of MAPK intervention. As heart failure, myocardial ischemia and arrhythmic dysfunction are associated with excessive local NE release, attenuation of NE release by H 3 R activation is cardioprotective. Thus, the uncovering of a novel H 3 R signaling pathway may ultimately bear therapeutic significance in hyper-adrenergic states.

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Section: Mapk Activation Plays a Role In The H 3 R-induced Inhibitionmentioning

confidence: 69%

Histamine H3-receptor signaling in cardiac sympathetic nerves: Identification of a novel MAPK-PLA2-COX-PGE2-EP3R pathway

Levi

Seyedi

Schäefer

et al. 2007

Biochemical Pharmacology

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“…The p38 inhibitor SB202190 does not a ect phosphorylation of p38 (data not shown) since the inhibitor only binds to the ATP pocket of p38 and blocks its intrinsic ATPase activity without a ecting its phosphorylation sites (Lee et al, 1994;Cuenda et al, 1995). In addition, the inhibition of p38 by SB202190 was reversible, therefore, the inhibitory e ect could not be detected by in vitro kinase assay of p38 (data not shown, Young et al, 1997;Wilson et al, 1997). Thus, we performed an in vivo kinase assay to determine p38 activity.…”

Section: Uvb Signi®cantly Activated P38 and Erkmentioning

confidence: 98%

Activation of p38 MAP kinase and ERK are required for ultraviolet-B induced c-fos gene expression in human keratinocytes

Chen

Bowden

1999

Oncogene

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The e ects of p38 MAP kinase and ERK on UVB induced c-fos gene expression were studied in a human keratinocyte cell line, FL30. UVB signi®cantly increased c-fos gene expression at both the transcriptional and protein levels. p38 and ERK were also signi®cantly activated after UVB irradiation. Treating the cells with p38 inhibitor SB202190 inhibited p38 activation, but not ERK; treating the cells with MEK-1 inhibitor PD98059 inhibited ERK activation without suppressing p38 activation. The kinase activation was determined by Western blots using phospho-p38 or ERK antibodies, or an in vivo p38 activity assay. Further studies demonstrated that blocking p38 almost completely abrogated UVB induced c-fos gene transcription and c-Fos protein synthesis. Inhibiting ERK partially abrogated UVB induced c-fos transcriptional and protein levels. Suppression of both p38 and ERK not only completely blocked UVB induced c-fos expression, but also decreased c-fos gene basal expression. Our data indicated that p38 may play a more important role than ERK in UVB induced cfos expression in human keratinocytes. Since c-fos expression may play an important role in UVB induced AP-1 activation, and AP-1 activation is known to play a role in tumor promotion, both p38 and ERK could be potential targets for chemoprevention of skin cancer.

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“…In order to obtain further evidence that the activation of c-Raf by SB 203580 results from its interaction with c-Raf, and not with SAPK2/p38, we transfected 293 cells with a SAPK2a/p38 mutant in which Thr 106 had been changed to Met to make it insensitive to low concentrations of SB 203580 (Wilson et al, 1997;Eyers et al, 1998). In cells transfected with this mutant, MAPKAP kinase-2 activity is elevated, but can no longer be suppressed by 1 mM SB 203580 in contrast to untransfected cells (Figure 7a).…”

Section: The Activation Of C-raf By Sb 203580 Is Not Caused By Interamentioning

confidence: 99%

“…SB 203580 binds competitively with ATP (Young et al, 1997) and the three-dimensional structure of SAPK2a/p38a in a complex with closely related pyridinyl imidazoles has established that these drugs are inserted into the ATP-binding pocket of SAPK2a/ p38a (Tong et al, 1997;Wilson et al, 1997). However, the 4-¯uorophenyl ring of the drug does not make contact with residues in the ATP-binding pocket that interact with ATP.…”

Section: Introductionmentioning

confidence: 99%

“…One residue that makes contact with the 4-¯uorophenyl moiety is Thr 106 and recent studies have established that this residue plays a critical role in determining sensitivity to SB 203580. Thus its mutation to Met or Gln, which are present at the equivalent position in other MAP kinase family members, or to other residues with bulky side chains, makes SAPK2a/p38a and SAPK2b/p38b2 insensitive to SB 203580 (Wilson et al, 1997;Eyers et al, 1998;Gum et al, 1998). Conversely, the mutation of this residue to Thr in other MAP kinase family members (SAPK1/JNK, SAPK3/p38g and SAPK4/p38d) makes them sensitive to SB 203580 (Eyers et al, 1998;Gum et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

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Effect of SB 203580 on the activity of c-Raf in vitro and in vivo

et al. 1999

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The inhibition of SAPK2a/p38 (a mitogen activated protein (MAP) kinase family member) by SB 203580 depends on the presence of threonine at residue 106. Nearly all other protein kinases are insensitive to this drug because a more bulky residue occupies this site (Eyers et al., 1998). Raf is one of the few protein kinases that possesses threonine at this position, and we show that SB 203580 inhibits c-Raf with an IC 50 of 2 mM in vitro. However, SB 203580 does not suppress either growth factor or phorbol ester-induced activation of the classical MAP kinase cascade in mammalian cells. One of the reasons for this is that SB 203580 also triggers a remarkable activation of c-Raf in vivo (when measured in the absence of the drug). The SB 203580-induced activation of c-Raf occurs without any increase in the GTP-loading of Ras, is not prevented by inhibitors of the MAPK cascade, protein kinase C or phosphatidylinositide 3-kinase, and is not triggered by the binding of this drug to SAPK2a/p38. The paradoxical activation of cRaf by SB 203580 (and by another structurally unrelated c-Raf inhibitor) suggests that inhibitors of the kinase activity of c-Raf may not be e ective as anti-cancer drugs.

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The structural basis for the specificity of pyridinylimidazole inhibitors of p38 MAP kinase

Abstract: Our results reveal how pyridinylimidazole compounds are potent and selective inhibitors of p38 MAP kinase but not other MAP kinases. It should now be possible to design other specific inhibitors of activated p38 MAP kinase using the structure of the nonphosphorylated enzyme.

Cited by 274 publications

References 31 publications

Histamine H3-receptor signaling in cardiac sympathetic nerves: Identification of a novel MAPK-PLA2-COX-PGE2-EP3R pathway

Histamine H3-receptor signaling in cardiac sympathetic nerves: Identification of a novel MAPK-PLA2-COX-PGE2-EP3R pathway

Activation of p38 MAP kinase and ERK are required for ultraviolet-B induced c-fos gene expression in human keratinocytes

Effect of SB 203580 on the activity of c-Raf in vitro and in vivo

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