Dolgikh D A, Kirpichnikov M P, Ptitsyn O B, Fedorov A N, Finkel'shteĭn A V, Chemeris V V
Mol Biol (Mosk). 1992 Nov-Dec;26(6):1242-50.
Based on the molecular theory of protein structure the de novo protein was designed in order to obtain the tertiary fold which has not yet been observed in natural proteins, namely four-stranded antiparallel beta-sheet covered by two alpha-helixes. The gene coding for this protein (named albebetin) was chemically synthesized, cloned in plasmid with SP6 phage promoter and expressed in mRNA-dependent cell-free translation system. An approach was developed to study albebetin using only nanogram amounts of radio labelled protein without previous purification. The preliminary analysis of its structure by gel-filtration, urea-gradient electrophoresis and limited proteolysis revealed compactness and stability of the de novo protein.
基于蛋白质结构的分子理论，设计了一种全新的蛋白质，以获得天然蛋白质中尚未观察到的三级折叠结构，即由两条α螺旋覆盖的四链反平行β折叠。编码这种蛋白质（命名为albebetin）的基因通过化学合成，克隆到带有SP6噬菌体启动子的质粒中，并在依赖mRNA的无细胞翻译系统中表达。开发了一种方法，仅使用纳克量的放射性标记蛋白质（无需预先纯化）来研究albebetin。通过凝胶过滤、尿素梯度电泳和有限蛋白酶解对其结构进行的初步分析表明，这种全新蛋白质具有紧密性和稳定性。
