Min Junhong, Kim Sang-Uk, Kim Young Jun, Yea Cheol-Heon, Choi Jeong-Woo
College of Bionanotechnology, Kyungwon University, Seognam-Si 461-701, Korea.
J Nanosci Nanotechnol. 2008 Oct;8(10):4982-7. doi: 10.1166/jnn.2008.1200.
A novel immobilization method of cupredoxin azurin on a gold surface was developed without a chemical linker using recombinant technique. A recombinant protein with cysteine residue by site-directed mutagenesis (SDM) was designed and then directly self-assembled on Au surface. The layer of the functionalized protein immobilized is confirmed by surface plasmon resonance (SPR) and its surface morphology is analyzed by scanning tunneling microscopy (STM). The immobilization efficiency has been increased about 73.2%, as compared to that of wild type azurin. The electrochemical property of the fabricated thin layer was investigated by the cyclic voltammetry (CV). As a result, cysteine-substituted azurin can be used for making high quality protein film, and applied to the fabrication of nano-scale bioelectronics.
利用重组技术，开发了一种无需化学连接体将铜蓝蛋白天青固定在金表面的新方法。通过定点诱变（SDM）设计了一种带有半胱氨酸残基的重组蛋白，然后直接在金表面进行自组装。通过表面等离子体共振（SPR）确认固定化功能化蛋白层，并通过扫描隧道显微镜（STM）分析其表面形态。与野生型天青相比，固定化效率提高了约73.2%。通过循环伏安法（CV）研究了制备的薄层的电化学性质。结果表明，半胱氨酸取代的天青可用于制备高质量的蛋白质膜，并应用于纳米级生物电子学的制造。
