PDBsum entry 3mh2

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Transferase PDB id
Jmol PyMol
Protein chain
330 a.a. *
Waters ×40
* Residue conservation analysis
PDB id:
Name: Transferase
Title: Mutagenesis of p38 map kinase establishes key roles of phe16 function and structural dynamics and reveals a novel dfg-ou
Structure: Mitogen-activated protein kinase 14. Chain: a. Synonym: map kinase 14, mapk 14, mitogen-activated protein alpha, map kinase p38 alpha, cytokine suppressive anti-infl drug-binding protein, csaid-binding protein, csbp, max-inte protein 2, map kinase mxi2, sapk2a. Engineered: yes. Mutation: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Gene: csbp, csbp1, csbp2, cspb1, mapk14, mxi2. Expressed in: escherichia coli. Expression_system_taxid: 562.
2.30Å     R-factor:   0.247     R-free:   0.318
Authors: H.V.Namboodiri,M.Karpusas,M.Bukhtiyarova,E.B.Springman
Key ref: M.Bukhtiyarova et al. (2007). Mutagenesis of p38alpha MAP kinase establishes key roles of Phe169 in function and structural dynamics and reveals a novel DFG-OUT state. Biochemistry, 46, 5687-5696. PubMed id: 17441692
07-Apr-10     Release date:   21-Apr-10    
Supersedes: 2pto
Go to PROCHECK summary

Protein chain
Pfam   ArchSchema ?
Q16539  (MK14_HUMAN) -  Mitogen-activated protein kinase 14
360 a.a.
330 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Enzyme reactions 
   Enzyme class: E.C.  - Mitogen-activated protein kinase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: ATP + a protein = ADP + a phosphoprotein
+ protein
+ phosphoprotein
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     extracellular region   12 terms 
  Biological process     intracellular signal transduction   65 terms 
  Biochemical function     nucleotide binding     13 terms  


Biochemistry 46:5687-5696 (2007)
PubMed id: 17441692  
Mutagenesis of p38alpha MAP kinase establishes key roles of Phe169 in function and structural dynamics and reveals a novel DFG-OUT state.
M.Bukhtiyarova, M.Karpusas, K.Northrop, H.V.Namboodiri, E.B.Springman.
In order to study the role of Phe169 in p38alpha MAP kinase structure and function, wild-type p38alpha and five p38alpha DFG motif mutants were examined in vitro for phosphorylation by MKK6, kinase activity toward ATF2 substrate, thermal stability, and X-ray crystal structure. All six p38alpha variants were efficiently phosphorylated by MKK6. However, only one activated p38alpha mutant (F169Y) possessed measurable kinase activity (1% compared to wild-type). The loss of kinase activity among the DFG mutants may result from an inability to correctly position Asp168 in the activated form of p38alpha. Two mutations significantly increased the thermal stability of p38alpha (F169A DeltaTm = 1.3 degrees C and D168G DeltaTm = 3.8 degrees C), and two mutations significantly decreased the stability of p38alpha (F169R DeltaTm = -3.2 degrees C and F169G DeltaTm = -4.7 degrees C). Interestingly, X-ray crystal structures of two thermally destabilized p38alpha-F169R and p38alpha-F169G mutants revealed a DFG-OUT conformation in the absence of an inhibitor molecule. This DFG-OUT conformation, termed alpha-DFG-OUT, is different from the ones previously identified in p38alpha crystal structures with bound inhibitors and postulated from high-temperature molecular dynamics simulations. Taken together, these results indicate that Phe169 is optimized for p38alpha functional activity and structural dynamics, rather than for structural stability. The alpha-DFG-OUT conformation observed for p38alpha-F169R and p38alpha-F169G may represent a naturally occurring intermediate state of p38alpha that provides access for binding of allosteric inhibitors. A model of the local forces driving the DFG IN-OUT transition in p38alpha is proposed.

Literature references that cite this PDB file's key reference

  PubMed id Reference
19109437 Y.Shan, M.A.Seeliger, M.P.Eastwood, F.Frank, H.Xu, M...Jensen, R.O.Dror, J.Kuriyan, and D.E.Shaw (2009).
A conserved protonation-dependent switch controls drug binding in the Abl kinase.
  Proc Natl Acad Sci U S A, 106, 139-144.  
18408713 A.Degterev, J.Hitomi, M.Germscheid, I.L.Ch'en, O.Korkina, X.Teng, D.Abbott, G.D.Cuny, C.Yuan, G.Wagner, S.M.Hedrick, S.A.Gerber, A.Lugovskoy, and J.Yuan (2008).
Identification of RIP1 kinase as a specific cellular target of necrostatins.
  Nat Chem Biol, 4, 313-321.  
18787129 A.P.Kornev, S.S.Taylor, and L.F.Ten Eyck (2008).
A helix scaffold for the assembly of active protein kinases.
  Proc Natl Acad Sci U S A, 105, 14377-14382.  
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