Expression, purification and crystallization of a human tau-tubulin kinase 2 that phosphorylates tau protein.
Tau-tubulin kinase 2 (TTBK2) is a Ser/Thr kinase that putatively phosphorylates residues Ser208 and Ser210 (numbered according to a 441-residue human tau isoform) in tau protein. Functional analyses revealed that a recombinant kinase domain (residues 1-331) of human TTBK2 expressed in insect cells with a baculovirus overexpression system retains kinase activity for tau protein. The kinase domain of TTBK2 was crystallized using the hanging-drop vapour-diffusion method. The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 55.6, b = 113.7, c = 117.3 A, alpha = beta = gamma = 90.0 degrees. Diffraction data were collected to 2.9 A resolution using synchrotron radiation at BL24XU of SPring-8.
Kitano-Takahashi
Michiko
M
Mitsubishi Kagaku Institute of Life Sciences, 11 Minamiooya, Machida, Tokyo 194-8511, Japan.
Morita
Hiroyuki
H
Kondo
Shin
S
Tomizawa
Kayoko
K
Kato
Ryohei
R
Tanio
Michikazu
M
Shirota
Yoshiko
Y
Takahashi
Hiroshi
H
Sugio
Shigetoshi
S
Kohno
Toshiyuki
T
D003460:Crystallization; D018360:Crystallography, X-Ray; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D016875:tau Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism
D016875:tau Proteins; C095903:tau-tubulin kinase; D017346:Protein Serine-Threonine Kinases