Kinetic Mechanism for p38 MAP Kinase

LoGrasso, Philip V.; Frantz, Betsy; Rolando, Anna M.; O’Keefe, Stephen J.; Hermes, Jeffery D.; O’Neill, Edward A.

doi:10.1021/bi9706778

Cited by 108 publications

(118 citation statements)

References 18 publications

(37 reference statements)

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“…P38-α induces, while p38-β suppresses cell death [36]. SB203580 inhibits mainly α and β isoforms [37,38]. Because no significant suppression of p38 was detected by SB203580, γ and/or δ p38 isoforms may be predominantly expressed in SVHK cells.…”

Section: Discussionmentioning

confidence: 99%

Differential phosphorylation of mitogen-activated protein kinase families by epidermal growth factor and ultraviolet B irradiation in SV40-transformed human keratinocytes

Takahashi

Kinouchi

Manabe

et al. 2001

Journal of Dermatological Science

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ELSEVIER, Nakamura, S; Takahashi, H; Kinouchi, M; Manabe, A; Ishida-Yamamoto, A; Hashimoto, Y; Iizuka, H, JOURNAL OF DERMATOLOGICAL SCIENCE, 25(2), 139-149, 2001. authorSV-40 transformed human keratinocytes (SVHK cells) were stimulated with epidermal growth factor (EGF)2 and ultraviolet B (UVB) irradiation. Following the stimulation, cell growth, apoptosis, and the activities of mitogen-activated protein (MAP) kinase families were analyzed. EGF (100ng/ml) increased SVHK cell number compared with control cells cultured in serum-free DMEM medium. The EGF-stimulated cells did not show DNA fragmentation. In contrast, UVB irradiation (40mJ/cm2) markedly decreased viable cell number, that was accompanied with DNA fragmentation. EGF stimulated extracellular signal-regulated kinase (ERK) and stress-activated protein kinase/c-Jun N-terminal kinase (JNK). Following the EGF stimulation, phosphorylated ERK and JNK were detected by phospho-p42/44 MAP kinase antibody and phospho-SAPK/JNK antibody, respectively. On the other hand, UVB irradiation stimulated the phosphorylation of p38 and JNK but not of ERK. The stimulation of ERK and JNK induced by EGF was observed earlier than the stimulation of p38 and JNK induced by UVB. PD98059, a specific MAP kinase kinase (MAPKK) 1 (also referred to as MEK1) inhibitor, inhibited EGF-dependent cell proliferation, that was associated with the inhibition of ERK and JNK phosphorylation. In contrast, UVB-induced overall cell death was not significantly affected by PD98059, that inhibited phosphorylation of JNK but not of p38. PD98059, however, significantly augmented UVB-induced cell death earlier time points (30min - 2 h). These results indicate that ERK and JNK are activated following EGF stimulation, that might be associated with cell proliferation. On the other hand, UVB-induced apoptosis seems to be mostly associated with the activation of p38. JNK stimulation might provide an anti-apoptotic tonus during the UVB-induced, p38-associated SVHK cell death

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Section: Discussionmentioning

confidence: 99%

Differential phosphorylation of mitogen-activated protein kinase families by epidermal growth factor and ultraviolet B irradiation in SV40-transformed human keratinocytes

Takahashi

Kinouchi

Manabe

et al. 2001

Journal of Dermatological Science

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“…For cAMPdependent kinase Whitehouse et al (50,51) reported that the mechanism was ordered, with the nucleotide binding first while Cook et al (30) reported that MgATP and peptide bind randomly, although initial binding of the nucleotide is preferred. An ordered sequential mechanism has been reported for p38 MAPK (32), for the vascular endothelial growth factor receptor-2 tyrosine kinase (52) and for the v-Src kinase (53). Both an ordered (54) and a random pathway (55) have been reported for the EGF receptor tyrosine kinase.…”

Section: Table II Summary Of Kinetic Constantsmentioning

confidence: 95%

The Cyclin-dependent Kinases cdk2 and cdk5 Act by a Random, Anticooperative Kinetic Mechanism

Clare¹,

Poorman²,

Kelley³

et al. 2001

Journal of Biological Chemistry

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cdk2⅐cyclin E and cdk5⅐p25 are two members of the cyclin-dependent kinase family that are potential therapeutic targets for oncology and Alzheimer's disease, respectively. In this study we have investigated the mechanism for these enzymes. Kinases catalyze the transfer of phosphate from ATP to a protein acceptor, thus utilizing two substrates, ATP and the target protein. For a two-substrate reaction, possible kinetic mechanisms include: ping-pong, sequential random, or sequential ordered. To determine the kinetic mechanism of cdk2⅐GST-cyclin E and cdk5⅐GST-p25, kinase activity was measured in experiments in which concentrations of peptide and ATP substrates were varied in the presence of dead-end inhibitors. A peptide identical to the peptide substrate, but with a substitution of valine for the phosphoacceptor threonine, competed with substrate with a K i value of 0.6 mM. An aminopyrimidine, PNU 112455A, was identified in a screen for inhibitors of cdk2. Nonlinear least squares and Lineweaver-Burk analyses demonstrated that the inhibitor PNU 112455A was competitive with ATP with a K i value of 2 M. In addition, a co-crystal of PNU 112455A with cdk2 showed that the inhibitor binds in the ATP binding pocket of the enzyme. Analysis of the inhibitor data demonstrated that both kinases use a sequential random mechanism, in which either ATP or peptide may bind first to the enzyme active site. For both kinases, the binding of the second substrate was shown to be anticooperative, in that the binding of the first substrate decreases the affinity of the second substrate. For cdk2⅐GST-cyclin E the kinetic parameters were determined to be K m, ATP ‫؍‬ 3.6 ؎ 1.0 M, K m, peptide ‫؍‬ 4.6 ؎ 1.4 M, and the anticooperativity factor, ␣ ‫؍‬ 130 ؎ 44. For cdk5⅐GST-p25, the K m, ATP ‫؍‬ 3.2 ؎ 0.7 M, K m, peptide ‫؍‬ 1.6 ؎ 0.3 M, and ␣ ‫؍‬ 7.2 ؎ 1.8.

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“…PD98059 23 (a specific inhibitor of ERK1/2), UO126 24 (a selective MEK inhibitor of ERK1/2 signaling) and SB203580 25 (a specific inhibitor of p38 MAPK) were added to the cells 1 hr before treatment with 100 lM EGCG plus 10 lM celecoxib: Pretreatments with PD98059 and UO126 dose-dependently reduced levels of phospho-ERK1/2 that were enhanced by cotreatment (Fig. 4a).…”

Section: Synergistic Induction Of Apoptosis By Cotreatment With Egcg mentioning

confidence: 99%

Green tea polyphenol stimulates cancer preventive effects of celecoxib in human lung cancer cells by upregulation of GADD153 gene

Suganuma

Kurusu

Suzuki

et al. 2006

Intl Journal of Cancer

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To more clearly understand the molecular mechanisms involved in synergistic enhancement of cancer preventive activity with the green tea polyphenol (2)-epigallocatechin gallate (EGCG), we examined the effects of cotreatment with EGCG plus celecoxib, a cyclooxygenase-2 selective inhibitor. We specifically looked for induction of apoptosis and expression of apoptosis related genes, with emphasis on growth arrest and DNA damage-inducible 153 (GADD153) gene, in human lung cancer cell line PC-9: Cotreatment with EGCG plus celecoxib strongly induced the expression of both GADD153 mRNA level and protein in PC-9 cells, while neither EGCG nor celecoxib alone did. However, cotreatment did not induce expression of other apoptosis related genes, p21 WAF1 and GADD45. Judging by upregulation of GADD153, only cotreatment with EGCG plus celecoxib synergistically induced apoptosis of PC-9 cells. Synergistic effects with the combination were also observed in 2 other lung cancer cell lines, A549 and ChaGo K-1. Furthermore, EGCG did not enhance GADD153 gene expression or apoptosis induction in PC-9 cells in combination with N-(4-hydroxyphenyl)retinamide or with aspirin. Thus, upregulation of GADD153 is closely correlated with synergistic enhancement of apoptosis with EGCG. Cotreatment also activated the mitogen-activated protein kinases (MAPKs), such as ERK1/2 and p38 MAPK: Preteatment with PD98059 (ERK1/2 inhibitor) and UO126 (selective MEK inhibitor) abrogated both upregulation of GADD153 and synergistic induction of apoptosis of PC-9 cells, while SB203580 (p38 MAPK inhibitor) did not do so, indicating that GADD153 expression was mediated through the ERK signaling pathway. These findings indicate that high upregulation of GADD153 is a key requirement for cancer prevention in combination with EGCG. ' 2006 Wiley-Liss, Inc.Key words: EGCG; apoptosis; ERK1/2; prevention (2)-Epigallocatechin gallate (EGCG), a main constituent of green tea polyphenol, can act synergistically with cancer preventive agents (such as sulindac and tamoxifen) in inducing apoptosis of lung, colon and breast cancer cells. 1-3 Since EGCG and green tea have a wide range of target organs and are nontoxic for animals and humans, there is growing interest in increasing the antitumor activity of cancer preventive agents in combination with EGCG or green tea. [4][5][6] However, the molecular mechanisms responsible for the synergistic effects of cotreatment with EGCG plus cancer preventive agents are not known. Using human cDNA cancer expression array, we have found upregulated expressions of 2 genes, growth arrest and DNA damage-inducible gene 153 (GADD153) and p21 WAF1 , and downregulated expressions of 4 genes, T-plasminogen activator, TIMP3, IL-1b and integrin b4: All of these genes are associated with synergistic induction of apoptosis of human non-small-cell lung carcinoma (NSCLC) cell line PC-9 by cotreatment with EGCG plus sulindac. 7 Treatment with either EGCG or sulindac alone did not affect these gene expressions. Since upregulation of GADD153 gene expressi...

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Kinetic Mechanism for p38 MAP Kinase

Cited by 108 publications

References 18 publications

Differential phosphorylation of mitogen-activated protein kinase families by epidermal growth factor and ultraviolet B irradiation in SV40-transformed human keratinocytes

Differential phosphorylation of mitogen-activated protein kinase families by epidermal growth factor and ultraviolet B irradiation in SV40-transformed human keratinocytes

The Cyclin-dependent Kinases cdk2 and cdk5 Act by a Random, Anticooperative Kinetic Mechanism

Green tea polyphenol stimulates cancer preventive effects of celecoxib in human lung cancer cells by upregulation of GADD153 gene

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