PDBsum entry 2baq

Transferase

PDB id: 2baq

Contents

Protein chain

330 a.a. *

Ligands

PQB

Waters ×7

* Residue conservation analysis

PDB id:

2baq

Name:

Transferase

Title:

P38alpha bound to ro3201195

Structure:

Mitogen-activated protein kinase 14. Chain: a. Synonym: map kinase p38 alpha. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: mapk14, csbp, csbp1, csbp2, mxi2. Expressed in: escherichia coli. Expression_system_taxid: 562. Expression_system_cell_line: bl21(de3).

Resolution:

2.80Å R-factor: 0.222 R-free: 0.296

Authors:

S.Gerhardt,R.A.Pauptit,J.Breed,J.Read,J.Tucker,R.A.Norman

Key ref:

J.E.Sullivan et al. (2005). Prevention of MKK6-dependent activation by binding to p38alpha MAP kinase. Biochemistry, 44, 16475-16490. PubMed id: 16342939 DOI: 10.1021/bi051714v

Date:

14-Oct-05 Release date: 06-Dec-05

Protein chain

Q16539  (MK14_HUMAN) -  Mitogen-activated protein kinase 14 from Homo sapiens

Seq: 360 a.a.

Struc: 330 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.2.7.11.24 - mitogen-activated protein kinase.
• Reaction:
  1. L-seryl-[protein] + ATP = O-phospho-L-seryl-[protein] + ADP + H⁺
  2. L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] + ADP + H⁺

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1021/bi051714v

Biochemistry 44:16475-16490 (2005)

PubMed id: 16342939

Prevention of MKK6-dependent activation by binding to p38alpha MAP kinase.

J.E.Sullivan, G.A.Holdgate, D.Campbell, D.Timms, S.Gerhardt, J.Breed, A.L.Breeze, A.Bermingham, R.A.Pauptit, R.A.Norman, K.J.Embrey, J.Read, W.S.VanScyoc, W.H.Ward.

ABSTRACT

Inhibition of p38alpha MAP kinase is a potential approach for the treatment of inflammatory disorders. MKK6-dependent phosphorylation on the activation loop of p38alpha increases its catalytic activity and affinity for ATP. An inhibitor, BIRB796, binds at a site used by the purine moiety of ATP and extends into a "selectivity pocket", which is not used by ATP. It displaces the Asp168-Phe169-Gly170 motif at the start of the activation loop, promoting a "DFG-out" conformation. Some other inhibitors bind only in the purine site, with p38alpha remaining in a "DFG-in" conformation. We now demonstrate that selectivity pocket compounds prevent MKK6-dependent activation of p38alpha in addition to inhibiting catalysis by activated p38alpha. Inhibitors using only the purine site do not prevent MKK6-dependent activation. We present kinetic analyses of seven inhibitors, whose crystal structures as complexes with p38alpha have been determined. This work includes four new crystal structures and a novel assay to measure K(d) for nonactivated p38alpha. Selectivity pocket compounds associate with p38alpha over 30-fold more slowly than purine site compounds, apparently due to low abundance of the DFG-out conformation. At concentrations that inhibit cellular production of an inflammatory cytokine, TNFalpha, selectivity pocket compounds decrease levels of phosphorylated p38alpha and beta. Stabilization of a DFG-out conformation appears to interfere with recognition of p38alpha as a substrate by MKK6. ATP competes less effectively for prevention of activation than for inhibition of catalysis. By binding to a different conformation of the enzyme, compounds that prevent activation offer an alternative approach to modulation of p38alpha.