PMCID
stringclasses 30
values | Title
stringclasses 30
values | Sentences
stringlengths 2
1.94k
|
|---|---|---|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Diffracting crystals for the phosporylation-mimicking CaMep2 DD mutant were obtained in space group P6322 from 0.1 M sodium acetate/15–20% PEG400, pH 5 (using decyl-maltoside as detergent; one molecule per AU), while S453D mutant crystals grew in 24% PEG400/0.05 M sodium acetate, pH 5.4/0.05 M magnesium acetate tetrahydrate/10 mM NH4Cl (space group R32; one molecule per AU).
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Finally, the 442Δ truncation mutant gave crystals under many different conditions, but most of these diffracted poorly or not at all.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
A reasonable low-resolution data set (3.4 Å resolution) was eventually obtained from a crystal grown in 24% PEG400/0.05 M sodium acetate/0.05 M magnesium acetate, pH 6.1 (space group R32).
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Diffraction data were collected at the Diamond Light Source and processed with XDS43 or HKL2000 (ref.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
44).
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
For MR, a search model was constructed with Sculptor within Phenix45, using a sequence alignment of ScMep2 with Archaeoglobus fulgidus Amt-1 (PDB ID 2B2H; ∼40% sequence identity to ScMep2).
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
A clear solution with nine molecules (three trimers) in the AU was obtained using Phaser46.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The model was subsequently completed by iterative rounds of manual building within Coot47 followed by refinement within Phenix.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The structures for WT CaMep2 were solved using the best-defined monomer of ScMep2 (60% sequence identity with CaMep2) in MR with Phaser, followed by automated model building within Phenix.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Finally, the structures of the three mutant CaMep2 proteins were solved using WT CaMep2 as the search model.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The data collection and refinement statistics for all six solved structures have been summarized in Supplementary Tables 1 and 2.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The S. cerevisiae haploid triple mepΔ strain (Σ1278b MATα mep1::LEU2 mep2::LEU2 mep3::G418 ura3-52) and triple mepΔ npr1Δ strain (Σ1278b MATα mep1::LEU2 mep2::LEU2 mep3::G418 npr1::NAT1 ura3-52) were generated by integrating the NAT1 resistance gene at one NPR1 locus in the diploid strain MLY131 (ref.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
6), followed by isolation of individual haploid strains.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Cells were grown in synthetic minimal medium with glucose (2%) as the carbon source and ammonium sulphate (1 mM) or glutamate (0.1%) as the nitrogen source.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Yeast cells were transformed as described48.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
All DNA sequences encoding epitope-tagged ScMep2 and its mutant derivatives were generated by PCR and homologous recombination using the vector pRS316 (ref.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
49).
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
In each case, the ScMEP2 sequences included the ScMEP2 promoter (1 kb), the ScMEP2 terminator and sequences coding for a His-6 epitope at the C-terminal end of the protein.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
All Mep2-His fusions contain the N4Q mutation to prevent glycosylation of Mep2 (ref.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
50).
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
All newly generated plasmid inserts were verified by DNA sequencing.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
For growth assays, S. cerevisiae cells containing plasmids expressing ScMep2 or mutant derivatives were grown overnight in synthetic minimal glutamate medium, washed, spotted by robot onto solid agar plates and culture growth followed by time course photography.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Images were then processed to quantify the growth of each strain over 3 days as described51.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The MODELLER (version 9.15) software package52 was used to build protein structures for MD simulations.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
This method was required to construct two complete protein models, the double mutant R452D/S453D (with the four missing residues from the X-ray structure added) and also the construct in which the mutation at position 452 is reverted to R, and D453 is replaced with a phosphoserine.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The quality of these models was assessed using normalized Discrete Optimized Protein Energy (DOPE) values and the molpdf assessment function within the MODELLER package.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The model R452D/S453D mutant has a molpdf assessment score of 1854.05, and a DOPE assessment score of -60920.55.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The model of the S453J mutant has a molpdf assessment score of 1857.01 and a DOPE assessment score of −61032.15.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
WT and model structures were embedded into a pre-equilibrated lipid bilayer composed of 512 dipalmitoylphosphatidylcholine lipids using the InflateGRO2 computer programme53.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The bilayers were then solvated with the SPC water model54 and counterions were added to achieve a charge neutral state.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
All simulations were performed with the GROMACS package (version 4.5.5)55, and the GROMOS96 43a1p force field.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
During simulation time, the temperature was maintained at 310 K using the Nosé-Hoover thermostat5657 with a coupling constant of 0.5 ps.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Pressure was maintained at 1 bar using semi-isotropic coupling with the Parrinello-Rahman barostat58 and a time constant of 5 ps.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Electrostatic interactions were treated using the smooth particle mesh Ewald algorithm59 with a short-range cutoff of 0.9 nm.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Van der Waals interactions were truncated at 1.4 nm with a long-range dispersion correction applied to energy and pressure.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The neighbour list was updated every five steps.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
All bonds were constrained with the LINCS algorithm60, so that a 2-fs time step could be applied throughout.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The phospholipid parameters for the dipalmitoylphosphatidylcholine lipids were based on the work of Berger61.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
The embedded proteins were simulated for 200 ns each; a repeat simulation was performed for each system with different initial velocities to ensure reproducibility.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
To keep the c.p.u.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
times within reasonable limits, all simulations were performed on Mep2 monomers.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
This is also consistent with previous simulations for E. coli AmtB161718.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Accession codes: The atomic coordinates and the associated structure factors have been deposited in the Protein Data Bank (http:// www.pdbe.org) with accession codes 5AEX (ScMep2), 5AEZ(CaMep2; R3), 5AF1(CaMep2; P3), 5AID(CaMep2; 442D), 5AH3 (CaMep2; R452D/S453D) and 5FUF (CaMep2; S453D).
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
How to cite this article: van den Berg, B. et al. Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Nat.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
Commun.
|
PMC4852598
|
Structural basis for Mep2 ammonium transceptor activation by phosphorylation.
|
7:11337 doi: 10.1038/ncomms11337 (2016).
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Mitogen-activated protein kinases (MAPKs), important in a large array of signalling pathways, are tightly controlled by a cascade of protein kinases and by MAPK phosphatases (MKPs).
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
MAPK signalling efficiency and specificity is modulated by protein–protein interactions between individual MAPKs and the docking motifs in cognate binding partners.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Two types of docking interactions have been identified: D-motif-mediated interaction and FXF-docking interaction.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Here we report the crystal structure of JNK1 bound to the catalytic domain of MKP7 at 2.4-Å resolution, providing high-resolution structural insight into the FXF-docking interaction.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The FNFL segment in MKP7 directly binds to a hydrophobic site on JNK1 that is near the MAPK insertion and helix αG. Biochemical studies further reveal that this highly conserved structural motif is present in all members of the MKP family, and the interaction mode is universal and critical for the MKP-MAPK recognition and biological function.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The mitogen-activated protein kinases (MAPKs) are central components of the signal-transduction pathways, which mediate the cellular response to a variety of extracellular stimuli, ranging from growth factors to environmental stresses123.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The MAPK signalling pathways are evolutionally highly conserved.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The basic assembly of MAPK pathways is a three-tier kinase module that establishes a sequential activation cascade: a MAPK kinase kinase activates a MAPK kinase, which in turn activates a MAPK.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The three best-characterized MAPK signalling pathways are mediated by the kinases extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The ERK pathway is activated by various mitogens and phorbol esters, whereas the JNK and p38 pathways are stimulated mainly by environmental stress and inflammatory cytokines456.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The MAPKs are activated by MAPK kinases that phosphorylate the MAPKs at conserved threonine and tyrosine residues within their activation loop.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
After activation, each MAPK phosphorylates a distinct set of protein substrates, which act as the critical effectors that enable cells to mount the appropriate responses to varied stimuli.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
MAPKs lie at the bottom of conserved three-component phosphorylation cascades and utilize docking interactions to link module components and bind substrates78.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Two types of docking motifs have been identified in MAPK substrates and cognate proteins: kinase-interacting motif (D-motif) and FXF-motif (also called DEF motif, docking site for ERK FXF).
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The best-studied docking interactions are those between MAP kinases and ‘D-motifs', which consists of two or more basic residues followed by a short linker and a cluster of hydrophobic residues.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The D-motif-docking site (D-site) in MAPKs is situated in a noncatalytic region opposite of the kinase catalytic pocket and is comprised of a highly acidic patch and a hydrophobic groove.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
D-motifs are found in many MAPK-interacting proteins, including substrates, activating kinases and inactivating phosphatases, as well as scaffolding proteins.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
A second docking motif for MAPKs consists of two Phe residues separated by one residue (FXF-motif).
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
This motif has been observed in several MAPK substrates910111213.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The FXF-motif-binding site of ERK2 has been mapped to a hydrophobic pocket formed between the P+1 site, αG helix and the MAPK insert14.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
However, the generality and mechanism of the FXF-mediated interaction is unclear.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The physiological outcome of MAPK signalling depends on both the magnitude and the duration of kinase activation15.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Downregulation of MAPK activity can be achieved through direct dephosphorylation of the phospho-threonine and/or tyrosine residues by various serine/threonine phosphatases, tyrosine phosphatases and dual-specificity phosphatases (DUSPs) termed MKPs.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
MKPs constitute a group of DUSPs that are characterized by their ability to dephosphorylate both phosphotyrosine and phosphoserine/phospho-threonine residues within a substrate1617.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Dysregulated expression of MKPs has been associated with pathogenesis of various diseases, and understanding their precise recognition mechanism presents an important challenge and opportunity for drug development1819.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Here, we present the crystal structure of JNK1 in complex with the catalytic domain of MKP7.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
This structure reveals the molecular mechanism underlying the docking interaction between MKP7 and JNK1.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
In the JNK1–MKP7 complex, a hydrophobic motif (FNFL) that initiates the helix α5 in the MKP7 catalytic domain directly binds to the FXF-motif-binding site on JNK1, providing the structural insight into the classic FXF-type docking interaction.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Biochemical and modelling studies further demonstrate that the molecular interactions mediate this key element for substrate recognition are highly conserved among all MKP-family members.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Thus, our study reveals a hitherto unrecognized interaction mode for encoding complex target specificity among MAPK isoforms.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
DUSPs belong to the protein-tyrosine phosphatases (PTPase) superfamily, which is defined by the PTPase-signature motif CXXGXXR20.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
MKPs represent a distinct subfamily within a larger group of DUSPs.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
In mammalian cells, the MKP subfamily includes 10 distinct catalytically active MKPs.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
All MKPs contain a highly conserved C-terminal catalytic domain (CD) and an N-terminal kinase-binding domain (KBD)1521.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The KBD is homologous to the rhodanese family and contains an intervening cluster of basic amino acids, which has been suggested to be important for interacting with the target MAPKs.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
On the basis of sequence similarity, substrate specificity and predominant subcellular localization, the MKP family can be further divided into three groups (Fig. 1).
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Biochemical and structural studies have revealed that the KBD of MKPs is critical for MKP3 docking to ERK2, and MKP5 binding to p38α, although their binding mechanisms are completely different2223.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
However, it is unknown if other MAPKs can interact with the KBD of their cognate phosphatases in the same manner as observed for recognition of ERK2 and p38α by their MKPs, or whether they recognize distinct docking motifs of MKPs.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
MKP7, the biggest molecule in the MKP family, selectively inactivates JNK and p38 following stress activation24.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
In addition to the CD and KBD, MKP7 has a long C-terminal region that contains both nuclear localization and export sequences by which MKP7 shuttles between the nucleus and the cytoplasm (Fig. 2a).
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
To quantitatively assess the contribution of the N-terminal domain to the MKP7-catalysed JNK1 dephosphorylation, we first measured the kinetic parameters of the C-terminal truncation of MKP7 (MKP7ΔC304, residues 5–303) and MKP7-CD (residues 156–301) towards phosphorylated JNK1 (pJNK1).
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Figure 2b shows the variation of initial rates of the MKP7ΔC304 and MKP7-CD-catalysed reaction with the concentration of phospho-JNK1.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Because the concentrations of MKP7 and pJNK1 were comparable in the reaction, the assumption that the free-substrate concentration is equal to the total substrate concentration is not valid.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Thus, the kinetic data were analysed using the general initial velocity equation, taking substrate depletion into account: The kcat and Km of the MKP7-CD (0.028 s and 0.26 μM) so determined were nearly identical to those of MKP7ΔC304 (0.029 s and 0.27 μM), indicating that the MKP7-KBD has no effect on enzyme catalysis.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
We next examined the interaction of JNK1 with the CD and KBD of MKP7 by gel filtration analysis.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
When 3 molar equivalents of CD were mixed with 1 molar equivalent of JNK1, a significant amount of CD co-migrated with JNK1 to earlier fractions, and the excess amount of CD was eluted from the size exclusion column as a monomer, indicating stable complex formation (Fig. 2c).
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
In contrast, no KBD–JNK1 complex was detected when 3 molar equivalents of KBD were mixed with 1 molar equivalent of JNK1.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
To further confirm the JNK1–MKP7-CD interaction, we performed a pull-down assay using the purified proteins.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
As shown in Fig. 2d, the CD of MKP7 can be pulled down by JNK1, while the KBD failed to bind to the counterpart protein.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
Taken together, our data indicate that the CD of MKP7, but not the KBD domain, is responsible for JNK substrate-binding and enzymatic specificity.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
To understand the molecular basis of JNK1 recognition by MKP7, we determined the crystal structure of unphosphorylated JNK1 in complex with the MKP7-CD (Fig. 3a, Supplementary Fig. 1a and Table 1).
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
In the complex, JNK1 has its characteristic bilobal structure comprising an N-terminal lobe rich in β-sheet and a C-terminal lobe that is mostly α-helical.
|
PMC4802042
|
A conserved motif in JNK/p38-specific MAPK phosphatases as a determinant for JNK1 recognition and inactivation.
|
The overall folding of MKP7-CD is typical of DUSPs, with a central twisted five-stranded β-sheet surrounded by six α-helices.
|
Subsets and Splits
No community queries yet
The top public SQL queries from the community will appear here once available.