The structure of phosphorylated p38gamma is monomeric and reveals a conserved activation-loop conformation.
BACKGROUND
Mitogen-activated protein (MAP) kinases mediate the cellular response to stimuli such as pro-inflammatory cytokines and environmental stress. P38gamma is a new member of the MAP kinase family, and is expressed at its highest levels in skeletal muscle. P38gamma is 63% identical in sequence to P38alpha. The structure of P38alpha MAP kinase has been determined in the apo, unphosphorylated, inactive form. The structures of apo unphosphorylated ERK2, a related MAP kinase, and apo phosphorylated ERK2 have also been determined.


RESULTS
We have determined the structure of doubly phosphorylated P38gamma in complex with an ATP analog by X-ray crystallography. This is the first report of a structure of an activated kinase in the P38 subfamily, and the first bound to a nucleotide. P38gamma residue phosphoryl-Thr183 forms hydrogen bonds with five basic amino acids, and these interactions induce an interdomain rotation. The conformation of the activation loop of P38gamma is almost identical to that observed in the structure of activated ERK2. However, unlike ERK2, the crystal structure and solution studies indicate that activated P38gamma exists as a monomer.


CONCLUSIONS
Interactions mediated by phosphoryl-Thr183 induce structural changes that direct the domains and active-site residues of P38gamma into a conformation consistent with catalytic activity. The conformation of the phosphorylation loop is likely to be similar in all activated MAP kinases, but not all activated MAP kinases form dimers.
Bellon
S
S
Vertex Pharmaceuticals Incorporated 130 Waverly Street, Cambridge, MA 02139-4211, USA. bellon@vpharm.com.
Fitzgibbon
M J
MJ
Fox
T
T
Hsiao
H M
HM
Wilson
K P
KP
D000266:Adenylyl Imidodiphosphate / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D018360:Crystallography, X-Ray; D019281:Dimerization; D004789:Enzyme Activation; D008274:Magnesium / Q000737:chemistry / Q000378:metabolism; D008715:Methionine / Q000737:chemistry / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000737:chemistry; D048057:Mitogen-Activated Protein Kinase 10; D020928:Mitogen-Activated Protein Kinases / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D010766:Phosphorylation; D011487:Protein Conformation; D011505:Protein-Tyrosine Kinases / Q000737:chemistry; D013912:Threonine / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases
D000266:Adenylyl Imidodiphosphate; D013912:Threonine; D008715:Methionine; D048057:Mitogen-Activated Protein Kinase 10; D011505:Protein-Tyrosine Kinases; D019950:Mitogen-Activated Protein Kinase 1; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D008274:Magnesium