Crystallization of MAP kinases.
X-ray structural studies of MAP kinases and MAP kinase module components are elucidating how kinase activity is regulated and how specificity of signaling is conferred. In the past decade, MAP kinases have been crystallized in their active, phosphorylated forms or low-activity, unphosphorylated forms, as well as in the presence of binding partners such as docking peptides and inhibitors. Crystallization has been achieved via diverse strategies including control of phosphorylation, coding sequence modification, incorporation of tags for purification, and use of a variety of cell-types for protein expression. Recently, interest has been focused on use of crystallography for lead optimization in the development for pharmacological inhibitors on MAP kinases. Further, some success has been gained in crystallizing the MAP kinase activators MAP2Ks and MAP3K kinase domains. This review describes the key methods that have been utilized to crystallize MAP kinases and MAP kinase pathway components.
Lee
Seung-Jae
SJ
Department of Biochemistry, The University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Boulevard, Dallas, TX 75390-8816, USA.
Zhou
Tianjun
T
Goldsmith
Elizabeth J
EJ
D000818:Animals; D001419:Bacteria / Q000235:genetics; D016367:Baculoviridae / Q000235:genetics; D003460:Crystallization / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism; D007313:Insecta / Q000166:cytology; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016296:Mutagenesis; D010766:Phosphorylation; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D020928:Mitogen-Activated Protein Kinases