Labutytė Greta, Povilonienė Simona, Šimoliūnas Eugenijus, Gabrielaitis Dovydas, Skapas Martynas, Noreika Algirdas, Meškys Rolandas, Časaitė Vida
Department of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Life Sciences Center, Vilnius University, Sauletekio av. 7, LT-10257 Vilnius, Lithuania.
Department of Neurobiology and Biophysics, Institute of Biosciences, Life Sciences Center, Vilnius University, Sauletekio av. 7, LT-10257 Vilnius, Lithuania.
Nanomaterials (Basel). 2021 Nov 12;11(11):3031. doi: 10.3390/nano11113031.
We report on the construction of functionalized nanotubes based on tail sheath protein 041 from vB_KleM-RaK2 bacteriophage. The truncated 041 protein (041Δ200) was fused with fluorescent proteins GFP and mCherry or amidohydrolase YqfB. The generated chimeric proteins were successfully synthesized in BL21 (DE3) cells and self-assembled into tubular structures. We detected the fluorescence of the structures, which was confirmed by stimulated emission depletion microscopy. When 041Δ200GFP and 041Δ200mCherry were coexpressed in BL21 (DE3) cells, the formed nanotubes generated Förster resonance energy transfer, indicating that both fluorescent proteins assemble into a single nanotube. Chimeric 041Δ200YqfB nanotubes possessed an enzymatic activity, which was confirmed by hydrolysis of -acetyl-2'-deoxycytidine. The enzymatic properties of 041Δ200YqfB were similar to those of a free wild-type YqfB. Hence, we conclude that 041-based chimeric nanotubes have the potential for the development of delivery vehicles and targeted imaging and are applicable as scaffolds for biocatalysts.
我们报道了基于来自vB_KleM-RaK2噬菌体的尾鞘蛋白041构建功能化纳米管的研究。截短的041蛋白（041Δ200）与荧光蛋白GFP和mCherry或酰胺水解酶YqfB融合。所产生的嵌合蛋白在BL21（DE3）细胞中成功合成，并自组装成管状结构。我们检测到了这些结构的荧光，这通过受激发射损耗显微镜得到了证实。当041Δ200GFP和041Δ200mCherry在BL21（DE3）细胞中共表达时，形成的纳米管产生了Förster共振能量转移，表明两种荧光蛋白都组装成了单个纳米管。嵌合的041Δ200YqfB纳米管具有酶活性，这通过对2'-脱氧胞苷的水解得到了证实。041Δ200YqfB的酶学性质与游离的野生型YqfB相似。因此，我们得出结论，基于041的嵌合纳米管具有开发递送载体和靶向成像的潜力，并且可作为生物催化剂的支架。
