Stabilization of Near-Infrared Fluorescent Proteins by Packaging in Virus-like Particles
Stabilization of Near-Infrared Fluorescent Proteins by Packaging in Virus-like Particles
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DOI：
10.1021/acs.biomac.0c00362
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发表时间：
2020-06-01
影响因子：
6.2
通讯作者：
Finn, M. G.
中科院分区：
文献类型：
--
作者：
Das, Soumen;Zhao, Liangjun;Finn, M. G.
近红外荧光Q β病毒样颗粒（VLPs）是通过在衣壳内包装高红移的胆汁素依赖荧光蛋白的单体和二聚体而高产出的。在大肠杆菌蛋白表达期间或之后，在培养基中简单添加盐酸胆绿素就足以产生完全成熟的封装荧光团。包装的近红外蛋白与未封装的类似物具有相同的光化学性质，但在热、混沌诱导变性和蛋白质水解方面更加稳定。无创体内成像显示，在小鼠全身注射后，VLPs主要运输到肝脏，表明这些颗粒很容易被标准仪器检测到。
Near-IR fluorescent Q beta virus-like particles (VLPs) were produced in a high yield by packaging highly red-shifted monomeric and dimeric versions of biliverdin-dependent fluorescent proteins within the capsid shell. The simple addition of biliverdin hydrochloride to the medium during or after Escherichia coli protein expression was enough to produce fully matured encapsidated fluorophores. The packaged near-IR proteins exhibited identical photochemical properties to their nonencapsidated analogues but were far more stable toward heat, chaotropeinduced denaturation, and proteolysis. Noninvasive in vivo imaging showed the VLPs to traffic primarily to the liver after systemic injection in mice, revealing that the particles were easily detected by a standard instrument.