Development of peptide-immobilized TiO2 chip for monitoring of intracellular protein phosphorylation signals.

研究了一种利用氧化钛板的肽阵列检测蛋白激酶活性的方法。首先，采用溶胶-凝胶法制备了氧化钛平板。该板具有相对平坦的表面和通过UV照射的光诱导表面润湿性转换的高亲水率。此外，我们成功地控制了表面润湿性可逆使用低压干燥和紫外线照射。其次，合成了聚丙烯酸与蛋白激酶A（PKA）底物肽的复合物（PAA/PEP）。我们用偶联酶法证实PAA/PEP被PKA磷酸化。最后，将荧光聚（丙烯酸）（F-PAA）固定在氧化钛板上，并将该板在水中超声处理。洗涤后，F-PAA仍然保留在板上。这表明F-PAA与钛氧化物板之间形成了结合。从以上结果，我们将建立基本的技术，以开发肽阵列使用氧化钛板。

A peptide array using a titanium oxide plate for detection of protein kinase activity has been investigated. First, the titanium oxide plate was prepared by sol-gel method. The plate has relatively flat surface and high hydrophilicity rate of photo-induced surface wettability conversion by UV irradiation. Furthermore, we succeeded that the surface wettability was controlled reversibly using hypobaric drying and UV irradiation. Next, complex with poly(acrylic)acid and protein kinase A (PKA) substrate peptide (PAA/PEP) was synthesized. We confirmed that PAA/PEP was phosphorylated by PKA, by coupled enzyme assay. Finally, fluorescenced poly(acrylic)acid (F-PAA) was immobilized on the titanium oxide plate, and the plate was sonicated in water. After washing, F-PAA still remained on the plate. It suggested that a bonding is formed between F-PAA and titanium oxide plate. From above results, we would establish the basis technique to develop the peptide array using the titanium oxide plate.