Dolgikh D A, Gabrielian A E, Uversky V N, Kirpichnikov M P
Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
Appl Biochem Biotechnol. 1996 Oct-Nov;61(1-2):85-96. doi: 10.1007/BF02785691.
The de novo protein albebetin has been engineered (J. Mol. Biol. 1992, 225, 927-931) to form a predesigned tertiary fold that has not yet been observed in natural proteins. Analysis of albebetin expressed in a cell-free system and in Escherichia coli revealed its compactness, relative stability, and the secondary structure close to the predesigned one. The blast-transforming biological activity of human interferon was grafted to albebetin by attachment of an eight amino acid interferon fragment to the N-terminus of albebetin next to its first methionine residue. The chimeric protein was expressed in a wheat germ cell-free translation system and tested for its structural properties, receptor binding, and biological activity. According to the tests, albebetin incorporating the active interferon fragment has a compact and relatively stable structure, and binds the murine thymocyte receptor effectively. It activates the blast transformation reaction of thymocyte cells even more efficiently than human interferon at low concentrations.
全新蛋白质阿尔贝贝汀已被设计改造（《分子生物学杂志》，1992年，第225卷，927 - 931页），以形成一种在天然蛋白质中尚未观察到的预先设计的三级结构。对在无细胞系统和大肠杆菌中表达的阿尔贝贝汀的分析揭示了其紧密性、相对稳定性以及与预先设计结构相近的二级结构。通过将一个八氨基酸的干扰素片段连接到阿尔贝贝汀第一个甲硫氨酸残基旁边的N端，将人干扰素的促细胞分裂转化生物活性嫁接到了阿尔贝贝汀上。该嵌合蛋白在小麦胚无细胞翻译系统中表达，并对其结构特性、受体结合和生物活性进行了测试。根据测试结果，掺入活性干扰素片段的阿尔贝贝汀具有紧密且相对稳定的结构，并能有效结合小鼠胸腺细胞受体。在低浓度下，它激活胸腺细胞的促细胞分裂转化反应甚至比人干扰素更有效。
