The structure of mitogen-activated protein kinase p38 at 2.1-Å resolution

Wang, Zhong Lin; Harkins, P.C.; Ulevitch, Richard J.; Han, Jiahuai; Cobb, Melanie H.; Goldsmith, Elizabeth J.

doi:10.1073/pnas.94.6.2327

Cited by 264 publications

(239 citation statements)

References 60 publications

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“…3A) used previously to identify domains important for interacting with upstream kinases, target substrates, and phosphatases (12,13,21). The three-dimensional structures of ERK1/2 and p38 are similar (22), and the chimeras contain selected intact structural subdomains from each enzyme. Binding to PEA-15 was assessed using lysates from CHO cells transfected with HA-tagged ERK/p38 chimeras incubated with immobilized GST-PEA-15 (Fig.…”

Section: Mutations Of Pea-15 That Block Erk1/2 Binding Abrogatementioning

confidence: 99%

PEA-15 Binding to ERK1/2 MAPKs Is Required for Its Modulation of Integrin Activation

Chou

Hill

Hsieh

et al. 2003

Journal of Biological Chemistry

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Activation of Raf-1 suppresses integrin activation, potentially through the activation of extracellular signalregulated kinases 1 and 2 (ERK1/2). However, bulk ERK1/2 activation does not correlate with suppression. PEA-15 reverses suppression of integrin activation and binds ERK1/2. Here we report that PEA-15 reversal of integrin suppression depends on its capacity to bind ERK1/2, indicating that ERK1/2 function is indeed required for suppression. Mutations in either the death effector domain or C-terminal tail of PEA-15 that block ERK1/2 binding abrogated the reversal of integrin suppression. Furthermore, we used ERK/p38 chimeras and site-directed mutagenesis to identify ERK1/2 residues required for binding PEA-15. Mutations of residues that precede the ␣G helix and within the mitogenactivated protein kinase insert blocked ERK2 binding to PEA-15, but not activation of ERK2. These ERK2 mutants blocked the ability of PEA-15 to reverse suppression of integrin activation. Thus, PEA-15 regulation of integrin activation depends on its binding to ERK1/2. To directly test the role of ERK1/2 localization in suppression, we enforced membrane association of ERK1 and 2 by joining a membrane-targeting CAAX box sequence to them. Both ERK1-CAAX and ERK2-CAAX were membrane-localized and suppressed integrin activation. In contrast to suppression by membrane-targeted Raf-CAAX, suppression by ERK1/2-CAAX was not reversed by PEA-15. Thus, ERK1/2 are the Raf effectors for suppression of integrin activation, and PEA-15 reverses suppression by binding ERK1/2.

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Section: Mutations Of Pea-15 That Block Erk1/2 Binding Abrogatementioning

confidence: 99%

PEA-15 Binding to ERK1/2 MAPKs Is Required for Its Modulation of Integrin Activation

Chou

Hill

Hsieh

et al. 2003

Journal of Biological Chemistry

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“…The crystal structures of unphosphorylated and phosphorylated ERK2 and unphosphorylated p38 have been solved (18,(22)(23)(24). Several important differences exist between the structures of the unphosphorylated forms of ERK2 and p38.…”

Section: Mitogen-activated Protein (Map)mentioning

confidence: 99%

“…The helix at the end of L16 is extended by 7 Å, and there is an increase in the hydrophilicity of residues that form contacts between L16 and the N-terminal domain. This results in a less intimate interaction between L16 and the N-terminal domain in p38 (22,24).Within MAP kinase cascades, the MEKs are the most specific enzymes. These dual specificity kinases activate their respective MAP kinase substrates by phosphorylating the threonine and tyrosine of the specific TXY sequence located in the phosphorylation lip.…”

mentioning

confidence: 99%

“…The MAP kinase insert and a loop consisting of residues 199 -205 of ERK2 interact with the phosphorylation lip in the unphosphorylated form of ERK2. Finally, a loop at the C terminus of the MAP kinases, L16 or the C-terminal tail, wraps around the back of the structure and interacts with the N-terminal domain (18).The crystal structures of unphosphorylated and phosphorylated ERK2 and unphosphorylated p38 have been solved (18,(22)(23)(24). Several important differences exist between the structures of the unphosphorylated forms of ERK2 and p38.…”

mentioning

confidence: 99%

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Phosphorylation of MAP Kinases by MAP/ERK Involves Multiple Regions of MAP Kinases

Wilsbacher¹,

Goldsmith

Cobb³

1999

Journal of Biological Chemistry

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Mitogen-activated protein (MAP) kinases are activated with great specificity by MAP/ERK kinases (MEKs). The basis for the specific activation is not understood. In this study chimeras composed of two MAP kinases, extracellular signal-regulated protein kinase 2 and p38, were assayed in vitro for phosphorylation and activation by different MEK isoforms to probe the requirements for productive interaction of MAP kinases with MEKs. Experimental results and modeling support the conclusion that the specificity of MEK/MAP kinase phosphorylation results from multiple contacts, including surfaces in both the N-and C-terminal domains. Mitogen-activated protein (MAP)1 kinase or extracellular signal-regulated protein kinase (ERK) cascades are present in all eukaryotes and are utilized in almost all signal transduction pathways originating from receptors at the cell surface (1, 2). A plethora of different stimuli, including growth factors, cytokines, heat shock, and ultraviolet light, can initiate signaling through these cascades. Each cascade consists of a three kinase module: a MAP kinase, a MAP kinase/ERK kinase (MEK) that activates the MAP kinase, and a MEK kinase (MEKK) that activates the MEK (3). The MAP kinase in each cascade preferentially phosphorylates substrates with a serine or threonine followed by a proline. There are three mammalian MAP kinase modules that have been extensively studied. These include the ERK1/2 module, the c-Jun N-terminal protein kinase/stressactivated protein kinase module, and the p38 module. ERK3, ERK4, and ERK5 and other p38 isoforms have also been identified, but the cascades leading to activation of these kinases are not well characterized (4 -11).Like other protein kinases, the MAP kinases are folded into two domains (12). The smaller N-terminal domain is composed mostly of ␤ strands, whereas the C-terminal domain is made up of ␣ helices. ATP binds between the two domains, and protein substrate is believed to bind on the surface of the C-terminal domain. Alignment of the amino acid sequences of many protein kinases reveals a common core catalytic domain of 250 -300 residues encoding the two-domain structure (13). Protein kinases possess 12 conserved stretches of amino acids within their catalytic domains known as subdomains (13-15). These conserved elements as well as unique structures contribute to catalysis by and regulation of protein kinases. The functions of several of the conserved and unique structural motifs are known. A glycine-rich loop in the N-terminal domain, termed the phosphate anchor ribbon, has a role in binding ATP. Also in the N-terminal domain, subdomain III encodes the C helix, which contains an invariant Glu involved in binding MgATP (16,17). This helix is important for maintaining an open domain conformation in unphosphorylated ERK2 (18), aligning catalytic residues in Src (19) and Cdk2 (20), and binding to the cyclin regulatory subunit in Cdk2 (21). The activation loop, known as the phosphorylation lip in MAP kinases, is a poorly conserved element in the C-termi...

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“…Among the X-ray structures of p38 MAP kinase (1IAN [14], 1WFC [15], 1P38 [16], among others), 1P38 was employed in the modeling for the following reasons. 1IAN forms a complex structure with 1 but only the Cα atom coordinates are known for the protein structure.…”

Section: Methodsmentioning

confidence: 99%

Complex Structure Modeling of p38 MAP Kinase and Pyridyl-pyrrole Compounds

Iwata

Nakao²,

Kagari

et al. 2001

CBIJ

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Some pyridyl-pyrrole compounds were found to have inhibitory activities against p38 MAP kinase. Their activity was much more potent than that of SB203580 that has a similar overall structure but with a different core group, i.e., an imidazole ring. The complex structure modeling was therefore carried out in an attempt to obtain insights into the interactions of the kinase with the pyridyl-pyrrole compounds and SB203580. The resultant complex models revealed that the imidazole and the pyrrole compounds had similar binding modes and that the representative interactions between the enzyme and the ligand moiety (pyridyl or phenyl) were common. For the sulfoxy moiety including the aliphatic tethers attached to a terminal functional group, the amino acid residues interacting with the moiety were elucidated. Lastly, the affinities of the inhibitors were estimated by calculating the score values with the Ludi program and a good correlation between the p38 MAP kinase inhibitory activities and the score values was found.

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The structure of mitogen-activated protein kinase p38 at 2.1-Å resolution

Cited by 264 publications

References 60 publications

PEA-15 Binding to ERK1/2 MAPKs Is Required for Its Modulation of Integrin Activation

PEA-15 Binding to ERK1/2 MAPKs Is Required for Its Modulation of Integrin Activation

Phosphorylation of MAP Kinases by MAP/ERK Involves Multiple Regions of MAP Kinases

Complex Structure Modeling of p38 MAP Kinase and Pyridyl-pyrrole Compounds

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