Engineering of a Red Fluorogenic Protein/Merocyanine Complex for Live-Cell Imaging.
Engineering of a Red Fluorogenic Protein/Merocyanine Complex for Live-Cell Imaging.
复制标题
用于活细胞成像的红色荧光蛋白/部花青复合物的工程。
DOI：
10.1002/cbic.201900428
复制
发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Borhan,Babak
中科院分区：
文献类型：
--
作者：
Santos,ElizabethM;Berbasova,Tetyana;Wang,Wenjing;Salmani,RaheleEsmatpour;Sheng,Wei;Vasileiou,Chrysoula;Geiger,JamesH;Borhan,Babak
重组人细胞视黄醇结合蛋白 II是一种15 kDa的蛋白，属于细胞内脂结合蛋白家族，它通过花青醛与活性中心赖氨酸残基的共价连接产生一种高荧光的红色色素。该复合体表现出“开启”的荧光，这是由于一种弱荧光的醛“发光”，随后在蛋白质的结合口袋内形成了一种强荧光的花青染料。花菁的细胞渗透速度很快，荧光团的成熟几乎是瞬间的。HCRBPII/花青素复合体具有高的量子产率、低的细胞毒性、细胞器的特异性标记以及与癌细胞和酵母细胞的相容性。HCRBPII/花青素标签比大多数常见的红色荧光蛋白更亮。
A reengineered human cellular retinol binding protein II (hCRBPII), a 15‐kDa protein belonging to the intracellular lipid binding protein (iLBP) family, generates a highly fluorescent red pigment through the covalent linkage of a merocyanine aldehyde to an active site lysine residue. The complex exhibits “turn‐on” fluorescence, due to a weakly fluorescent aldehyde that “lights up” with subsequent formation of a strongly fluorescent merocyanine dye within the binding pocket of the protein. Cellular penetration of merocyanine is rapid, and fluorophore maturation is nearly instantaneous. The hCRBPII/merocyanine complex displays high quantum yield, low cytotoxicity, specificity in labeling organelles, and compatibility in both cancer cell lines and yeast cells. The hCRBPII/merocyanine tag is brighter than most common red fluorescent proteins.