Expression, purification and crystallization of a human tau-tubulin kinase 2 that phosphorylates tau protein

Abstract

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.

Figure 1

Coomassie Brilliant Blue-stained SDS–PAGE gel (10–20% gradient) of the purified recombinant human TTBK2 (1–331). Lane 1, molecular-weight markers; lane 2, human TTBK2 (1–331).

Figure 2

Crystal of TTBK2 (1–331). The dimensions of the crystals were approximately 0.15 × 0.08 × 0.08 mm.