scholarly journals Structural Studies of Soybean Calmodulin Isoform 4 Bound to the Calmodulin-binding Domain of Tobacco Mitogen-activated Protein Kinase Phosphatase-1 Provide Insights into a Sequential Target Binding Mode

2009 ◽  
Vol 284 (41) ◽  
pp. 28292-28305 ◽  
Author(s):  
Hiroaki Ishida ◽  
Mario Rainaldi ◽  
Hans J. Vogel
Keyword(s):  
Protein Kinase ◽  
Binding Mode ◽  
Binding Domain ◽  
Mitogen Activated Protein Kinase ◽  
Structural Studies ◽  
Calmodulin Binding ◽  
Mitogen Activated Protein ◽  
Target Binding

Structure of mitogen-activated protein kinase kinase 1 in the DFG-out conformation

2021 ◽  
Vol 77 (12) ◽  
Author(s):  
Setsu Nakae ◽  
Maho Kitamura ◽  
Daisuke Fujiwara ◽  
Masaaki Sawa ◽  
Tsuyoshi Shirai ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Protein Kinase ◽  
Binding Mode ◽  
Mitogen Activated Protein Kinase ◽  
Structural Elements ◽  
Atp Binding Site ◽  
Starting Point ◽  
Mitogen Activated Protein ◽  
Activation Segment ◽  
Protein Kinase Kinase

Eukaryotic protein kinases contain an Asp-Phe-Gly (DFG) motif, the conformation of which is involved in controlling the catalytic activity, at the N-terminus of the activation segment. The motif can be switched between active-state (DFG-in) and inactive-state (DFG-out) conformations: however, the mechanism of conformational change is poorly understood, partly because there are few reports of the DFG-out conformation. Here, a novel crystal structure of nonphosphorylated human mitogen-activated protein kinase kinase 1 (MEK1; amino acids 38–381) complexed with ATP-γS is reported in which MEK1 adopts the DFG-out conformation. The crystal structure revealed that the structural elements (the αC helix and HRD motif) surrounding the active site are involved in the formation/stabilization of the DFG-out conformation. The ATP-γS molecule was bound to the canonical ATP-binding site in a different binding mode that has never been found in previously determined crystal structures of MEK1. This novel ATP-γS binding mode provides a starting point for the design of high-affinity inhibitors of nonphosphorylated inactive MEK1 that adopts the DFG-out conformation.

scholarly journals Oxidative stress inhibits MEKK1 by site-specific glutathionylation in the ATP-binding domain

2004 ◽  
Vol 381 (3) ◽  
pp. 675-683 ◽  
Author(s):  
Janet V. CROSS ◽  
Dennis J. TEMPLETON
Keyword(s):  
Oxidative Stress ◽  
Protein Kinase ◽  
Cysteine Residue ◽  
Binding Domain ◽  
Mitogen Activated Protein Kinase ◽  
Atp Binding ◽  
Cysteine Oxidation ◽  
Mitogen Activated Protein ◽  
In Cells

Many intracellular signalling events are accompanied by generation of reactive oxygen species in cells. Oxidation of protein thiol groups is an emerging theme in signal-transduction research. We have found that MEKK1 [MAPK (mitogen-activated protein kinase)/ERK (extracellular-signal-regulated kinase) kinase kinase 1], an upstream activator of the SAPK/JNK (stress-activated protein kinase/c-Jun N-terminal kinase) pathway, is directly inhibited by cysteine alkylation using NEM (N-ethylmaleimide). The related kinase, ASK1 (apoptosis signal-regulating kinase 1), was not inhibited, but was instead activated by NEM. Inhibition of MEKK1 requires a single unique cysteine residue (Cys1238) in the ATP-binding domain of MEKK1. Oxidative stress induced by menadione (2-methyl-1,4-naphthoquinone) also inhibited MEKK1, but activated ASK1, in cells. MEKK1 inhibition by menadione also required Cys1238. Oxidant-inhibited MEKK1 was re-activated by dithiothreitol and glutathione, supporting reversible cysteine oxidation as a mechanism. Using various chemical probes, we excluded modification by S-nitrosylation or oxidation of cysteine to sulphenic acid. Oxidant-inhibited MEKK1 migrated normally on non-reducing gels, excluding the possibility of intra- or inter-molecular disulphide bond formation. MEKK1 was inhibited by glutathionylation in vitro, and MEKK1 isolated from menadione-treated cells was shown by MS to be modified by glutathione on Cys1238. Our results support a model whereby the redox environment within the cell selectively regulates stress signalling through MEKK1 versus ASK1, and may thereby participate in the induction of apoptosis by oxidative stress.

scholarly journals Unique Signaling Properties of CTAR1 in LMP1-Mediated Transformation

2007 ◽  
Vol 81 (18) ◽  
pp. 9680-9692 ◽  
Author(s):  
Bernardo A. Mainou ◽  
David N. Everly ◽  
Nancy Raab-Traub
Keyword(s):  
Cell Cycle ◽  
Protein Kinase ◽  
Signaling Pathway ◽  
Tumor Necrosis Factor Receptor ◽  
Binding Domain ◽  
Dominant Negative ◽  
Mitogen Activated Protein Kinase ◽  
Barr Virus ◽  
Mitogen Activated Protein ◽  
Epstein Barr

ABSTRACT The Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) gene is considered the EBV oncogene as it is necessary for EBV-mediated transformation of B lymphocytes and itself transforms rodent fibroblasts. LMP1 activates the NF-κB, phosphatidylinositol 3-kinase (PI3K)-Akt, mitogen-activated protein kinase, and Jun N-terminal protein kinase signaling pathways through its two signaling domains, carboxyl-terminal activating regions 1 and 2 (CTAR1 and CTAR2). CTAR1 and CTAR2 induce signal transduction pathways through their direct (CTAR1) or indirect (CTAR2) recruitment of tumor necrosis factor receptor-associated factors (TRAFs). CTAR1 is necessary for LMP1-mediated transformation as well as activation of PI3K signaling and induction of cell cycle markers associated with G1/S transition. In this study, activation of PI3K-Akt signaling and deregulation of cell cycle markers were mapped to the TRAF-binding domain within CTAR1 and to the residues between CTAR1 and CTAR2. LMP1 CTAR1 also activated the MEK1/2-extracellular signal-regulated kinase 1/2 signaling pathway, and this activation was necessary for LMP1-induced transformation of Rat-1 fibroblasts. Dominant-negative forms of TRAF2 and TRAF3 inhibited but did not fully block LMP1-mediated transformation. These findings identify a new signaling pathway that is uniquely activated by the TRAF-binding domain of LMP1 and is required for transformation.

scholarly journals 1H, 15N and 13C sequence specific backbone assignment of the MAP Kinase Binding Domain of the Dual Specificity Phosphatase 1 and its interaction with the MAPK p38

2020 ◽  
Author(s):  
Ganesan Senthil Kumar ◽  
Rebecca Page ◽  
Wolfgang Peti
Keyword(s):  
Protein Kinase ◽  
Map Kinase ◽  
Backbone Assignment ◽  
Binding Domain ◽  
Mitogen Activated Protein Kinase ◽  
Dual Specificity Phosphatase ◽  
Associated Diseases ◽  
Dual Specificity ◽  
Mitogen Activated Protein

AbstractThe sequence-specific backbone assignment of the mitogen-activated protein kinase (MAPK) binding domain of the dual-specificity phosphatase 1 (DUSP1) has been accomplished using a uniformly [13C,15N]-labeled protein. These assignments will facilitate further studies of DUSP1 in the presence of inhibitors/ligands to target MAPK associated diseases and provide further insights into the function of dual-specificity phosphatase 1 in MAPK regulation.

Sign in / Sign up

Export Citation Format

Share Document