Role of a cysteine residue in the active site of ERK and the MAPKK family

Ohori, Makoto; Kinoshita, Takayoshi; Yoshimura, Seiji; Warizaya, Masaichi; Nakajima, Hidenori; Miyake, Hiroshi

doi:10.1016/j.bbrc.2006.12.083

Cited by 79 publications

(73 citation statements)

References 12 publications

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“…Such binding has the potential to alter or inhibit protein structure and function. A review of the literature identified a reactive cysteine residue within the ATP binding domain of MEK, C166, that is a target for Michael addition by other small electrophilic compounds, including FR148083 (35). The ATP binding domain of ERK is highly conserved with that of MEK and contains a cysteine residue in the same position (35).…”

Section: Discussionmentioning

confidence: 99%

“…A review of the literature identified a reactive cysteine residue within the ATP binding domain of MEK, C166, that is a target for Michael addition by other small electrophilic compounds, including FR148083 (35). The ATP binding domain of ERK is highly conserved with that of MEK and contains a cysteine residue in the same position (35). Here, we demonstrate that biotinylated 15d-PGJ 2 interacts directly with MEK and ERK and that preincubation with FR148083 inhibits binding of 15d-PGJ 2 to MEK ( Figure 7D).…”

Section: Discussionmentioning

confidence: 99%

“…As expected, biotin 15d-PGJ 2 did not interact with p38 (data not shown). A review of the literature identified a putative binding site for small electrophilic compounds in the ATP binding domain of MEK and ERK and a commercially available small molecule inhibitor, FR148083, known to target this binding site (35). We therefore pretreated HLFs with FR148083 before adding biotin15d-PGJ 2 , and associated proteins were precipitated as described above.…”

Section: D-pgj 2 Directly Interacts With Mek and Erkmentioning

confidence: 99%

See 2 more Smart Citations

Inhibition of Transglutaminase 2, a Novel Target for Pulmonary Fibrosis, by Two Small Electrophilic Molecules

Olsen

Epa²,

Kulkarni

et al. 2014

Am J Respir Cell Mol Biol

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Idiopathic pulmonary fibrosis (IPF) is characterized by progressive fibrotic destruction of normal lung architecture. Due to a lack of effective treatment options, new treatment approaches are needed. We previously identified transglutaminase (TG)2, a multifunctional protein expressed by human lung fibroblasts (HLFs), as a positive driver of fibrosis. TG2 catalyzes crosslinking of extracellular matrix proteins, enhances cell binding to fibronectin and integrin, and promotes fibronectin expression. We investigated whether the small electrophilic molecules 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid (CDDO) and 15-deoxy-delta-12,14-prostaglandin J 2 (15d-PGJ 2 ) inhibit the expression and profibrotic functions of TG2. CDDO and 15d-PGJ 2 reduced expression of TG2 mRNA and protein in primary HLFs from control donors and donors with IPF. CDDO and 15d-PGJ 2 also decreased the in vitro profibrotic effector functions of HLFs including collagen gel contraction and cell migration. The decrease in TG2 expression did not occur through activation of the peroxisome proliferator activated receptor g or generation of reactive oxidative species. CDDO and 15d-PGJ 2 inhibited the extracellular signal-regulated kinase pathway, resulting in the suppression of TG2 expression. This is the first study to show that small electrophilic compounds inhibit the expression and profibrotic effector functions of TG2, a key promoter of fibrosis. These studies identify new and important antifibrotic activities of these two small molecules, which could lead to new treatments for fibrotic lung disease.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: D-pgj 2 Directly Interacts With Mek and Erkmentioning

confidence: 99%

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Inhibition of Transglutaminase 2, a Novel Target for Pulmonary Fibrosis, by Two Small Electrophilic Molecules

Olsen

Epa²,

Kulkarni

et al. 2014

Am J Respir Cell Mol Biol

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“…f152A1 was reported previously as LL-Z1640-2 (Ellestad et al, 1978) or 5Z-7-oxo-zeaenol (Rawlins et al, 1999). The biological activities of f152A1 and its derivatives have been reported: inhibition of growth and motility of ciliated protozoan T. pyriformis (Ellestad et al, 1978); inhibition of IL-1␤ release from human monocytes (Kastelic et al, at ASPET Journals on May 11, 2018 jpet.aspetjournals.org Downloaded from 1996); inhibition of endotoxin-induced TNF␣ production from U937 cells (Rawlins et al, 1999); inhibition of T-cell activation (Camacho et al, 1999), Ras signaling (Tanaka et al, 1999), JNK/p38 pathways (Takehana et al, 1999), and TAK1 activation (Ninomiya-Tsuji et al, 2003); and inhibition of MEK (Zhao et al, 1999;Ohori et al, 2007). The number of reported in vivo effects of f152A1 and related compounds is more limited.…”

Section: Discussionmentioning

confidence: 99%

E6201 [(3S,4R,5Z,8S,9S,11E)-14-(Ethylamino)-8, 9,16-trihydroxy-3,4-dimethyl-3,4,9,19-tetrahydro-1H-2-benzoxacyclotetradecine-1,7(8H)-dione], a Novel Kinase Inhibitor of Mitogen-Activated Protein Kinase/Extracellular Signal-Regulated Kinase Kinase (MEK)-1 and MEK Kinase-1: In Vitro Characterization of Its Anti-Inflammatory and Antihyperproliferative Activities

Goto¹,

Chow²,

Muramoto³

et al. 2009

J Pharmacol Exp Ther

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The goal of this study is to identify a novel inhibitor with antiinflammatory and antiproliferative properties for the treatment of psoriasis. Compound f152A1 [(3S,5Z,8S,11E)-8,9,16-trihydroxy-14-methoxy-3-methyl-3,4,9,10-tetrahydro-1H-benzo[c][1] oxacyclotetradecine1,7(8H)-dione] was identified as the main active metabolite with strong inhibitory activity against tumor necrosis factor-␣ (TNF␣) transcription in a fraction originated from the fermentation broth of the fungus Curvularia verruculosa. Although active in cell-based assays, f152A1 was unstable in plasma and liver microsome preparations, thus limiting its pharmaceutical utilization. To improve the metabolic properties of f152A1, a medicinal chemistry program was undertaken, resulting in the generation of over 400 analogs of f152A1. Eventually, E6201 [(3S,4R,5Z,8S,9S,11E)-14-(ethylamino)-8, 9,16-trihydroxy-3,4-dimethyl-3,4,9,19-tetrahydro-1H-2-benzoxacyclotetradecine-1,7(8H)-dione] was identified as a promising analog in this series. In the present study, we characterized the in vitro activities of E6201 and discovered that the compound inhibits lipopolysaccharide-activated TNF␣ reporter activity in THP-1-33 cells with an IC 50 value of 50 nM and selectively inhibits mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK)-1 and MEK kinase-1 in cell-free biochemical assays. In addition, E6201 showed inhibitory activity in several other cell-based systems: 1) phosphorylation of c-jun N-terminal kinase and p38 MAPKs; 2) nuclear factor-B and activated protein-1 activation in various cell types; 3) interleukin (IL)-2 production from human lymphocytes; 4) hyperproliferation of human keratinocytes; 5) IL-8 production from human keratinocytes; and 6) proinflammatory cytokine production from human peripheral blood mononuclear cells. Based on the data presented here, E6201 may be beneficial for treatment of inflammatory and hyperproliferative diseases such as psoriasis through its anti-inflammatory activities on immune cells and antihyperproliferative activities on keratinocytes.This work was supported by Eisai Co., Ltd. Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.

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“…Therefore, it is likely that oxidation of Cys196 may interfere with the Mg-ATP binding function of D197, leading to the inactivation of the kinase. Similarly, in the case of ERK that also has a cysteine (i.e., Cys 166 ) immediately in front of the DFG triplet, covalent modification of Cys 166 by FR148083, a small compound isolated from fermentation culture broth, led to specific inactivation of ERK (35). Both JNK and p38 MAPK were resistant to FR148083 due to the lack of such a cysteine in front of the DFG triplet.…”

Section: Discussionmentioning

confidence: 99%

Oxidation-induced intramolecular disulfide bond inactivates mitogen-activated protein kinase kinase 6 by inhibiting ATP binding

Diao

Liu

Wong

et al. 2010

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

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Mitogen-activated protein kinase kinase 6 (MKK6) is a member of the mitogen-activated protein kinase (MAPK) kinase (MAP2K) subfamily that specifically phosphorylates and activates the p38 MAPKs. Based on both biochemical and cellular assays, we found that MKK6 was extremely sensitive to oxidation: It was inactivated by oxidation and its kinase activity was fully restored upon treatment with a reducing agent. Detailed mechanistic studies showed that cysteines 109 and 196, two of the six cysteines in MKK6, formed an intramolecular disulfide bond upon oxidation that inactivated MKK6 by inhibiting its ATP binding. This mechanism is distinct from that seen in other redox-sensitive kinases. The two cysteines involved in intramolecular disulfide formation are conserved in all seven members of the MAP2K family. Consistently, we confirmed that other MAP2Ks were also sensitive to oxidation. Our work reveals that MKK6 and other MAP2Ks are a distinct class of cellular redox sensors.

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Role of a cysteine residue in the active site of ERK and the MAPKK family

Cited by 79 publications

References 12 publications

Inhibition of Transglutaminase 2, a Novel Target for Pulmonary Fibrosis, by Two Small Electrophilic Molecules

Inhibition of Transglutaminase 2, a Novel Target for Pulmonary Fibrosis, by Two Small Electrophilic Molecules

Oxidation-induced intramolecular disulfide bond inactivates mitogen-activated protein kinase kinase 6 by inhibiting ATP binding

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