De novo design of functional metalloproteins

2015 Fiscal Year Final Research Report

• Project/Area Number: 25410174
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Nagoya Institute of Technology
• Principal Investigator: tanaka tosiki 名古屋工業大学, 工学(系)研究科(研究院), 教授 (70171775)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: デノボタンパク質 / 金属タンパク質 / 銅タンパク質 / 銅配位構造 / EPR測定

Outline of Final Research Achievements

Natural proteins are usually unstable toward heat and in organic solvents. We introduced couple of disulfide bonds in designed 3-helical bundled proteins. The protein was stable at 90° and in 50% EtOH. As for the design of cupper configuration, we constructed the noble copper configuration using two His and two Cys residues. Hemocyanin has two Cu ions bridged by two oxygens. This site has not constructed in the de novo designed helical proteins. Based on the knowledge obtained in blue copper protein, we utilized a Cys residue for a transient copper binding site, and succeeded to place the two His residues in nearby site. However, bridging by two oxygens was not attained so far. In conclusion, we succeeded in construction of the quite stable scaffold protein structure, noble copper configuration, and two copper proteins in vicinity. Theses techniques are quite promising to construct noble functional proteins, that does not exist in natural proteins, in near future.

Free Research Field

タンパク質工学