Activity of the MAP kinase ERK2 is controlled by a flexible surface loop

Jiandong Zhang, Faming Zhang, Douglas Ebert, Melanie H. Cobb, Elizabeth J. Goldsmith

Research output: Contribution to journalArticle

106 Citations (Scopus)

Abstract

Background: The mitogen-activated protein (MAP) kinase, ERK2, is a tightly regulated enzyme in the ubiquitous Ras-activated protein kinase cascade. ERK2 is activated by phosphorylation at two sites, Y185 and T183, that lie in the phosphorylation lip at the mouth of the catalytic site. To ascertain the role of these two residues in securing the low-activity conformation of the enzymes we have carried out crystallographic analyses and assays of phosphorylation-site mutants of ERK2. Results The crystal structures of four mutants, T183E (threonine at residue 183 is replaced by glutamate), Y185E, Y185F and the double mutant T183E/Y185E, were determined. When T183 is replaced by glutamate, few conformational changes are observed. By contrast, when Y185 is replaced by glutamate, 19 residues become disordered, including the entire phosphorylation lip and an adjacent loop. The conservative substitution of phenylalanine for Y185 also induces relatively large conformational changes. A binding site for phosphotyrosine in the active enzyme is putatively identified on the basis of the high-resolution refinement of the structure of wild-type ERK2. Conclusion The remarkable disorder observed throughout the phosphorylation lip when Y185 is mutated shows that the stability of the phosphorylation lip is rather low. Therefore, only modest amounts of binding energy will be required to dislodge the lip for phosphorylation, and it is likely that these residues will be involved in conformational changes associated both with binding to kinases and phosphatases and with activation. Furthermore, the low-activity structure is specifically dependent on Y185, whereas there is no such dependency on T183. Both residues, however, participate in forming the active enzyme, contributing to its tight control.

Original languageEnglish (US)
Pages (from-to)299-307
Number of pages9
JournalStructure
Volume3
Issue number3
DOIs
StatePublished - 1995

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Mitogen-Activated Protein Kinases
Phosphorylation
Lip
Glutamic Acid
Enzymes
ras Proteins
Phosphotyrosine
Threonine
Phenylalanine
Phosphoric Monoester Hydrolases
Protein Kinases
Mouth
Catalytic Domain
Phosphotransferases
Binding Sites

Keywords

  • crystallography
  • MAP kinase ERK2
  • MEK
  • phosphorylation sites

ASJC Scopus subject areas

  • Molecular Biology
  • Structural Biology

Cite this

Activity of the MAP kinase ERK2 is controlled by a flexible surface loop. / Zhang, Jiandong; Zhang, Faming; Ebert, Douglas; Cobb, Melanie H.; Goldsmith, Elizabeth J.

In: Structure, Vol. 3, No. 3, 1995, p. 299-307.

Research output: Contribution to journalArticle

Zhang, Jiandong ; Zhang, Faming ; Ebert, Douglas ; Cobb, Melanie H. ; Goldsmith, Elizabeth J. / Activity of the MAP kinase ERK2 is controlled by a flexible surface loop. In: Structure. 1995 ; Vol. 3, No. 3. pp. 299-307.
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