Crystallization and preliminary X-ray analysis of two inhibitor complexes of the catalytic domain of death-associated protein kinase.
The catalytic domain of death-associated protein kinase (DAPK) has been overexpressed, purified and crystallized using the sitting-drop vapour-diffusion method with PEG 8000 and magnesium acetate as precipitants. Complexes with the inhibitor staurosporine and its analogue BDB402 were also crystallized in the presence of PEG 400 and PEG 8000, respectively. Diffraction data were collected to 2.4 A for the native catalytic domain, to 2.9 A for the staurosporine complex and to 2.7 A for the BDB402 complex. All three crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 77.992, b = 109.909, c = 50.063 A for the catalytic domain, a = 78.911, b = 113.162, c = 50.658 A for the staurosporine complex and a = 77.337, b = 108.869, c = 50.186 A for the BDB402 complex. In both complexes the inhibitor molecule was clearly assigned in the difference Fourier map calculated on the basis of the phases obtained from the structure of the catalytic domain.
Yamakawa
Akira
A
Devision of Chemistry, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
Ogata
Hideaki
H
Morita
Kumiko
K
Shibata
Naoki
N
Andou
Naoko
N
Sanuki
Hiromi
H
Yamada
Kazunori
K
Hioki
Takeshi
T
Ishii
Takehisa
T
Higuchi
Yoshiki
Y
D000818:Animals; D051017:Apoptosis Regulatory Proteins; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D020134:Catalytic Domain; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D064550:Death-Associated Protein Kinases; D004791:Enzyme Inhibitors / Q000737:chemistry*; D006801:Humans; D019311:Staurosporine / Q000737:chemistry
D051017:Apoptosis Regulatory Proteins; D004791:Enzyme Inhibitors; D064550:Death-Associated Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D019311:Staurosporine