Catalytically active MAP KAP kinase 2 structures in complex with staurosporine and ADP reveal differences with the autoinhibited enzyme.
MAP KAP kinase 2 (MK2), a Ser/Thr kinase, plays a crucial role in the inflammatory process. We have determined the crystal structures of a catalytically active C-terminal deletion form of human MK2, residues 41-364, in complex with staurosporine at 2.7 A and with ADP at 3.2 A, revealing overall structural similarity with other Ser/Thr kinases. Kinetic analysis reveals that the K(m) for ATP is very similar for MK2 41-364 and p38-activated MK2 41-400. Conversely, the catalytic rate and binding for peptide substrate are dramatically reduced in MK2 41-364. However, phosphorylation of MK2 41-364 by p38 restores the V(max) and K(m) for peptide substrate to values comparable to those seen in p38-activated MK2 41-400, suggesting a mechanism for regulation of enzyme activity.
Underwood
Kathryn W
KW
Department of Biological Chemistry, Wyeth Research, 87 Cambridge Park Drive, Cambridge, MA 02140, USA. kunderwood@wyeth.com
Parris
Kevin D
KD
Federico
Elizabeth
E
Mosyak
Lidia
L
Czerwinski
Robert M
RM
Shane
Tania
T
Taylor
Meggin
M
Svenson
Kristine
K
Liu
Yan
Y
Hsiao
Chu-Lai
CL
Wolfrom
Scott
S
Maguire
Michelle
M
Malakian
Karl
K
Telliez
Jean-Baptiste
JB
Lin
Lih-Ling
LL
Kriz
Ronald W
RW
Seehra
Jasbir
J
Somers
William S
WS
Stahl
Mark L
ML
D000244:Adenosine Diphosphate / Q000378:metabolism*; D000595:Amino Acid Sequence; D004789:Enzyme Activation; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D046911:Macromolecular Substances; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D019311:Staurosporine / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases; D000099190:MAP-Kinase-Activated Kinase 2
D000244:Adenosine Diphosphate; D047908:Intracellular Signaling Peptides and Proteins; D046911:Macromolecular Substances; D020928:Mitogen-Activated Protein Kinases; D017346:Protein Serine-Threonine Kinases; D019311:Staurosporine; D048051:p38 Mitogen-Activated Protein Kinases; D000099190:MAP-Kinase-Activated Kinase 2