Department of Physical and Environmental Sciences, University of Toronto at Scarborough, Toronto, Canada.
Analyst. 2012 May 7;137(9):2042-6. doi: 10.1039/c2an35097a. Epub 2012 Mar 23.
Hyperphosphorylation of Tau, a protein that stabilizes microtubules, leads to the breakdown of the microtubular structure and ultimately to the formation of neurofibrillar tangles within neurons. Here, we report monitoring of Tau phosphorylations electrochemically, using Tau protein films chemically linked to gold surfaces and 5'-γ-ferrocenyl (Fc) adenosine triphosphate (Fc-ATP) as a co-substrate. Fc-phosphorylation reactions of Tau are explored using the three protein kinases, glycogen synthase kinase (GSK-3β), sarcoma (Src)-related kinase, and protein kinase A (PKA), which catalyze Fc-phosphorylation of different residues and regions within Tau. The kinetic parameters of the biochemical process (K(M) and V(max)) were determined.
tau 蛋白的过度磷酸化会导致微管结构的破坏,最终导致神经元内神经原纤维缠结的形成。在这里,我们报告了使用 tau 蛋白膜的电化学监测,该蛋白膜通过化学方式与金表面连接,并使用 5'-γ- 二茂铁 (Fc) 腺苷三磷酸 (Fc-ATP) 作为共底物。使用三种蛋白激酶——糖原合酶激酶 (GSK-3β)、肉瘤 (Src) 相关激酶和蛋白激酶 A (PKA)——探索了 Tau 的 Fc 磷酸化反应,这些激酶催化 Tau 内不同残基和区域的 Fc 磷酸化。确定了生化过程的动力学参数 (K (M) 和 V (max))。