Bender Gretchen M, Lehmann Andreas, Zou Hongling, Cheng Hong, Fry H Christopher, Engel Don, Therien Michael J, Blasie J Kent, Roder Heinrich, Saven Jeffrey G, DeGrado William F
Department of Biochemistry and Molecular Biophysics, Johnson Foundation, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
J Am Chem Soc. 2007 Sep 5;129(35):10732-40. doi: 10.1021/ja071199j. Epub 2007 Aug 10.
We describe the computational design of a single-chain four-helix bundle that noncovalently self-assembles with fully synthetic non-natural porphyrin cofactors. With this strategy, both the electronic structure of the cofactor as well as its protein environment may be varied to explore and modulate the functional and photophysical properties of the assembly. Solution characterization (NMR, UV-vis) of the protein showed that it bound with high specificity to the desired cofactors, suggesting that a uniquely structured protein and well-defined site had indeed been created. This provides a genetically expressed single-chain protein scaffold that will allow highly facile, flexible, and asymmetric variations to enable selective incorporation of different cofactors, surface-immobilization, and introduction of spectroscopic probes.
我们描述了一种单链四螺旋束的计算设计,该单链四螺旋束与完全合成的非天然卟啉辅因子非共价自组装。通过这种策略,可以改变辅因子的电子结构及其蛋白质环境,以探索和调节组装体的功能和光物理性质。对该蛋白质的溶液表征(核磁共振、紫外可见光谱)表明,它与所需的辅因子具有高特异性结合,这表明确实构建了一种结构独特的蛋白质和明确的位点。这提供了一种基因表达的单链蛋白质支架,它将允许高度简便、灵活和不对称的变化,以实现不同辅因子的选择性掺入、表面固定以及光谱探针的引入。