Bioengineering of Halobacterium sp. NRC-1 gas vesicle nanoparticles with GvpC fusion protein produced in E. coli.
Bioengineering of Halobacterium sp. NRC-1 gas vesicle nanoparticles with GvpC fusion protein produced in E. coli.
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DOI：
10.1007/s00253-022-11841-1
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发表时间：
2022-03
影响因子：
5
通讯作者：
DasSarma S
中科院分区：
文献类型：
--
作者：
Kim JM;Kim YS;Kim YR;Choi MJ;DasSarma P;DasSarma S
气体囊泡纳米颗粒（GVNPs）是用于细胞浮选的中空、浮力的原核细胞器。GVNPs由盐古菌（Halobacterium sp. NRC-1）中的一个大气体囊泡蛋白（gvp）基因簇编码，其中包括一个基因gvpC，该基因指定了与纳米颗粒表面结合的蛋白质。通过将外源序列与gvpC融合，在盐杆菌中产生了基因工程GVNPs。为了提高GvpC平台的通用性，我们开发了一种在盐古菌纳米颗粒上显示外源生产的GvpC融合蛋白的方法。在GvpC的c端或c端附近融合了链球菌igg结合蛋白结构域，从大肠杆菌中表达和纯化，并显示与野生型GVNPs结合。GvpC3GB和GvpC4GB两种融合蛋白，无论是否具有高酸性GvpC c端区，都能很好地结合纳米颗粒。gvnp结合的gvpc - igg结合融合蛋白也能够与酶连接的IgG-HRP复合物结合，并保持酶活性，证明杂交系统具有展示和传递蛋白质复合物的能力。这是第一个证明外源生产的GvpC融合蛋白与野生型盐古菌GvpC功能结合的报告，这极大地扩展了平台生产生物医学应用的生物工程纳米颗粒的能力。•卤古菌气体囊泡纳米颗粒（GVNPs）构成了一个多功能的显示系统。•大肠杆菌中表达的gvppc -链球菌igg结合融合蛋白与GVNPs结合。•在浮动GVNPs上显示的igg结合蛋白结合并显示IgG-HRP复合物。
Gas vesicle nanoparticles (GVNPs) are hollow, buoyant prokaryotic organelles used for cell flotation. GVNPs are encoded by a large gas vesicle protein (gvp) gene cluster in the haloarchaeon, Halobacterium sp. NRC-1, including one gene, gvpC, specifying a protein bound to the surface of the nanoparticles. Genetically engineered GVNPs in the Halobacterium sp. have been produced by fusion of foreign sequences to gvpC. To improve the versatility of the GVNP platform, we developed a method for displaying exogenously produced GvpC fusion proteins on the haloarchaeal nanoparticles. The streptococcal IgG-binding protein domain was fused at or near the C-terminus of GvpC, expressed and purified from E. coli, and shown to bind to wild-type GVNPs. The two fusion proteins, GvpC3GB and GvpC4GB, without or with a highly acidic GvpC C-terminal region, were found to be able to bind nanoparticles equally well. The GVNP-bound GvpC-IgG-binding fusion protein was also capable of binding to an enzyme-linked IgG-HRP complex which retained enzyme activity, demonstrating the hybrid system capability for display and delivery of protein complexes. This is the first report demonstrating functional binding of exogenously produced GvpC fusion proteins to wild-type haloarchaeal GVNPs which significantly expands the capability of the platform to produce bioengineered nanoparticles for biomedical applications. • Haloarchaeal gas vesicle nanoparticles (GVNPs) constitute a versatile display system. • GvpC-streptococcal IgG-binding fusion proteins expressed in E. coli bind to GVNPs. • IgG-binding proteins displayed on floating GVNPs bind and display IgG-HRP complex.
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DOI：
10.1016/s2213-2600(21)00559-2
发表时间：
2022-03
期刊：
The Lancet. Respiratory medicine
影响因子：
--
作者：
Burki TK
通讯作者：
Burki TK
影响因子：
7.8
作者：
DasSarma S;DasSarma P
通讯作者：
DasSarma P
影响因子：
4.6
作者：
Balakrishnan, Arjun;DasSarma, Priya;Chakravortty, Dipshikha
通讯作者：
Chakravortty, Dipshikha
影响因子：
5.5
作者：
Childs, Tawanna S.;Webley, Wilmore C.
通讯作者：
Webley, Wilmore C.
影响因子：
3
作者：
Dutta, Sneha;DasSarma, Priya;Jarori, Gotam K.
通讯作者：
Jarori, Gotam K.