Presentation is loading. Please wait.

Presentation is loading. Please wait.

Catalytic Activation of the Phosphatase MKP-3 by ERK2 Mitogen-Activated Protein Kinase by Montserrat Camps, Anthony Nichols, Corine Gillieron, Bruno Antonsson,

Published byDavid Fedde de Vos Modified over 5 years ago

Similar presentations

Presentation on theme: "Catalytic Activation of the Phosphatase MKP-3 by ERK2 Mitogen-Activated Protein Kinase by Montserrat Camps, Anthony Nichols, Corine Gillieron, Bruno Antonsson,"— Presentation transcript:

1 Catalytic Activation of the Phosphatase MKP-3 by ERK2 Mitogen-Activated Protein Kinase
by Montserrat Camps, Anthony Nichols, Corine Gillieron, Bruno Antonsson, Marco Muda, Christian Chabert, Ursula Boschert, and Steve Arkinstall Science Volume 280(5367): May 22, 1998 ©1998 by American Association for the Advancement of Science

2 Figure 1 Activation of MKP-3 by ERK2.
Activation of MKP-3 by ERK2. Phosphatase activity was measured as p-NPP hydrolysis at 25°C monitored at an absorbance of 405 nm (A405) (10). Full-length GST–MKP-3 and GST-ERK2 were expressed inE. coli and purified on glutathione-Sepharose. ERK2 was further purified by ion-exchange chromatography (9,10). (A) Time-dependent hydrolysis ofp-NPP by 5 μg of GST–MKP-3 either alone (0 μg) or in the presence of the indicated amounts of ERK2. Linear reaction rates are indicated by lines of best fit. Ten micrograms of ERK2 stimulated phosphatase activity by 25.5-fold. (B) Activation of the indicated amounts of GST–MKP-3 by purified ERK2. Incubations were for 30 min. Maximal ERK2-dependent activation with 10 μg of MKP-3 was 35 times the basal phosphatase activity. Data points are the mean of duplicate determinations and are representative of three independent experiments. Montserrat Camps et al. Science 1998;280: ©1998 by American Association for the Advancement of Science

3 Figure 2 Requirement of the MKP-3 catalytic Cys-293 and binding to the MKP-3 NH2-terminus for ERK2-dependent activation. Requirement of the MKP-3 catalytic Cys-293 and binding to the MKP-3 NH2-terminus for ERK2-dependent activation. Phosphatase activity was measured as described (Fig. 1). Incubations were performed with the indicated concentrations of (A) WT GST–MKP-3, (B) inactive mutant GST–MKP-3 C293S, or (C) NH2-terminally truncated GST–MKP-3ΔN (amino acids 153 to 381) in the absence (○) or presence (•) of ERK2 (5 μg). (D) Inhibition of ERK2-dependent activation of MKP-3 by the indicated concentrations of mutant inactive GST–MKP-3 (C293S) (▿), the MKP-3 NH2-terminus GST-MKP-3ΔC (amino acids 1 to 221) (•), or GST protein (○) incubated in the presence of GST–MKP-3 (2.5 μg) and ERK2 (2.5 μg). (▪) MKP-3 activity in the absence of ERK2. Data represent the mean of two identical experiments each performed in triplicate. Montserrat Camps et al. Science 1998;280: ©1998 by American Association for the Advancement of Science

4 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

5 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

6 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

7 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

8 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

9 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

10 Figure 5 Resistance of ERK2 D319N Sevenmaker to inactivation by MKP-3.
Resistance of ERK2 D319N Sevenmaker to inactivation by MKP-3. Wild-type ERK2 (A and C) or ERK2 D319N (B) (0.5 μg) were incubated in vitro in the absence (lane 1) or presence (all other lanes) of 0.1 μg of constitutively active MEK1 (S217E S221E) (16) and with the indicated concentrations (0.01 to 10 μg) of full-length MKP-3 (A and B) or MKP-3ΔN (amino acids 153 to 381) (C). Figure shows autoradiograms of MBP phosphorylation by activated MAP kinases. Experiments were repeated three times with identical results. Montserrat Camps et al. Science 1998;280: ©1998 by American Association for the Advancement of Science

11 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

12 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

13 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

14 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

15 Montserrat Camps et al. Science 1998;280:1262-1265
©1998 by American Association for the Advancement of Science

Download ppt "Catalytic Activation of the Phosphatase MKP-3 by ERK2 Mitogen-Activated Protein Kinase by Montserrat Camps, Anthony Nichols, Corine Gillieron, Bruno Antonsson,"

Similar presentations

Volume 13, Issue 2, Pages (February 2011)

Volume 13, Issue 2, Pages (February 2011)
Figure 1. Enhancement of ERα activity by Constitutively Active MEKK1 in Endometrial Cancer Cells A, Ishikawa cells were transfected with 0.5 μg EREe1bCAT.

Figure 1. Enhancement of ERα activity by Constitutively Active MEKK1 in Endometrial Cancer Cells A, Ishikawa cells were transfected with 0.5 μg EREe1bCAT.
RNA aptamers as pathway-specific MAP kinase inhibitors

RNA aptamers as pathway-specific MAP kinase inhibitors
Volume 91, Issue 7, Pages (December 1997)

Volume 91, Issue 7, Pages (December 1997)
by Chong-Shan Shi, Joseph Tuscano, and John H. Kehrl

by Chong-Shan Shi, Joseph Tuscano, and John H. Kehrl
Volume 13, Issue 2, Pages (January 2004)

Volume 13, Issue 2, Pages (January 2004)
TEL-JAK2 constitutively activates the extracellular signal–regulated kinase (ERK), stress-activated protein/Jun kinase (SAPK/JNK), and p38 signaling pathways.

TEL-JAK2 constitutively activates the extracellular signal–regulated kinase (ERK), stress-activated protein/Jun kinase (SAPK/JNK), and p38 signaling pathways.
A Cytoplasmic Inhibitor of the JNK Signal Transduction Pathway

A Cytoplasmic Inhibitor of the JNK Signal Transduction Pathway
Phosphorylation of NF-κB p65 by PKA Stimulates Transcriptional Activity by Promoting a Novel Bivalent Interaction with the Coactivator CBP/p300  Haihong.

Phosphorylation of NF-κB p65 by PKA Stimulates Transcriptional Activity by Promoting a Novel Bivalent Interaction with the Coactivator CBP/p300  Haihong.
Volume 3, Issue 1, Pages (January 1999)

Volume 3, Issue 1, Pages (January 1999)
Activating and Silencing the Mitotic Checkpoint through CENP-E-Dependent Activation/Inactivation of BubR1  Yinghui Mao, Ariane Abrieu, Don W. Cleveland 

Activating and Silencing the Mitotic Checkpoint through CENP-E-Dependent Activation/Inactivation of BubR1  Yinghui Mao, Ariane Abrieu, Don W. Cleveland 
Volume 87, Issue 7, Pages (December 1996)

Volume 87, Issue 7, Pages (December 1996)
Volume 8, Issue 16, Pages (July 1998)

Volume 8, Issue 16, Pages (July 1998)
Direct Activation of Gastric H,K-ATPase by N-Terminal Protein Kinase C Phosphorylation. Comparison of the Acute Regulation Mechanisms of H,K-ATPase and.

Direct Activation of Gastric H,K-ATPase by N-Terminal Protein Kinase C Phosphorylation. Comparison of the Acute Regulation Mechanisms of H,K-ATPase and.
Neurexins Are Functional α-Latrotoxin Receptors

Neurexins Are Functional α-Latrotoxin Receptors
The Molecular Basis of Aichi Virus 3A Protein Activation of Phosphatidylinositol 4 Kinase IIIβ, PI4KB, through ACBD3  Jacob A. McPhail, Erik H. Ottosen,

The Molecular Basis of Aichi Virus 3A Protein Activation of Phosphatidylinositol 4 Kinase IIIβ, PI4KB, through ACBD3  Jacob A. McPhail, Erik H. Ottosen,
X-Ray Crystal Structure and Functional Analysis of Vaccinia Virus K3L Reveals Molecular Determinants for PKR Subversion and Substrate Recognition  Arvin.

X-Ray Crystal Structure and Functional Analysis of Vaccinia Virus K3L Reveals Molecular Determinants for PKR Subversion and Substrate Recognition  Arvin.
Phosphorylation of PML by mitogen-activated protein kinases plays a key role in arsenic trioxide-mediated apoptosis  Fumihiko Hayakawa, Martin L Privalsky 

Phosphorylation of PML by mitogen-activated protein kinases plays a key role in arsenic trioxide-mediated apoptosis  Fumihiko Hayakawa, Martin L Privalsky 
Volume 13, Issue 2, Pages (February 2011)

Volume 13, Issue 2, Pages (February 2011)
Volume 91, Issue 2, Pages (October 1997)

Volume 91, Issue 2, Pages (October 1997)

Similar presentations

Ads by Google