Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United States.
Biomacromolecules. 2011 Nov 14;12(11):3977-81. doi: 10.1021/bm200983k. Epub 2011 Oct 13.
Qβ virus-like particles encapsulating multiple copies of fluorescent proteins were generated in high yields using a modular system enhanced by specific engineered RNA--protein interactions. The resulting particles were structurally indistinguishable from recombinant Qβ alone. The encapsidated proteins were nearly identical in photochemical properties to monomeric analogues, were more stable toward thermal degradation, and were protected from proteolytic cleavage. Residues on the outer capsid surface were chemically derivatized by acylation and azide--alkyne cycloaddition without affecting the fluorescence properties of the packaged proteins. A high-affinity carbohydrate-based ligand of the CD22 receptor was thereby attached, and specific cell labeling by the particles was successfully detected by flow cytometry and confocal laser microscopy.
利用增强特定工程化 RNA-蛋白相互作用的模块化系统,以高产率生成了包封多个荧光蛋白拷贝的 Qβ 病毒样颗粒。所得颗粒在结构上与单独的重组 Qβ 无法区分。包封的蛋白质在光化学性质上与单体类似物几乎相同,对热降解更稳定,并且免受蛋白水解切割的影响。通过酰化和叠氮化物-炔烃环加成反应对外壳表面的残基进行化学衍生化,而不会影响包装蛋白的荧光性质。随后,将 CD22 受体的高亲和力基于碳水化合物的配体连接上,并通过流式细胞术和共聚焦激光显微镜成功检测到颗粒的特异性细胞标记。