Visualization of molecular interactions using bimolecular fluorescence complementation analysis: characteristics of protein fragment complementation.
Visualization of molecular interactions using bimolecular fluorescence complementation analysis: characteristics of protein fragment complementation.
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DOI：
10.1039/b909638h
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发表时间：
2009-10
影响因子：
46.2
通讯作者：
Kerppola TK
中科院分区：
文献类型：
--
作者：
Kerppola TK
对维持生命的分子过程的研究必须包括对正常细胞环境中这些过程的研究。双分子荧光互补(BIFC)分析为显示活细胞中蛋白质的相互作用和修饰提供了一种方法。这种分析是基于与相互作用伙伴融合的荧光蛋白的互补片段的促进关联。相互作用伙伴形成的复合体将荧光蛋白片段连接在一起，这可以促进它们的结合。BIFC分析能够以高空间分辨率对蛋白质复合体进行敏感的可视化。BIFC分析的时间分辨率受到荧光团形成所需时间的限制，以及通过结合荧光蛋白片段来稳定复合体的时间。已经开发了许多对BIFC试验的修改和增强。多色BIFC分析可以同时显示同一细胞中的多个蛋白质复合体，并可用于研究细胞内相互排斥的相互作用伙伴之间对复合体形成的竞争。泛素介导的荧光互补(UbFC)分析方法可以显示细胞中的共价泛素家族多肽与底物蛋白的结合。BIFC分析也可以用来观察蛋白质与细胞中特定的染色质结构域和其他分子支架的结合。因此，BIFC分析为可视化影响活细胞中分子接近的各种过程提供了一种强有力的方法。
Investigation of the molecular processes that sustain life must include studies of these processes in the normal cellular environment. The bimolecular fluorescence complementation (BiFC) assay provides an approach for the visualization of protein interactions and modifications in living cells. This assay is based on the facilitated association of complementary fragments of a fluorescent protein that are fused to interaction partners. Complex formation by the interaction partners tethers the fluorescent protein fragments in proximity to each other, which can facilitate their association. The BiFC assay enables sensitive visualization of protein complexes with high spatial resolution. The temporal resolution of BiFC analysis is limited by the time required for fluorophore formation as well as the stabilization of complexes by association of the fluorescent protein fragments. Many modifications and enhancements of the BiFC assay have been developed. The multicolor BiFC assay enables simultaneous visualization of multiple protein complexes in the same cell and can be used to investigate the competition among mutually exclusive interaction partners for complex formation in cells. The ubiquitin-mediated fluorescence complementation (UbFC) assay enables visualization of covalent ubiquitin family peptide conjugation to substrate proteins in cells. The BiFC assay can also be used to visualize protein binding to specific chromatin domains as well as other molecular scaffolds in cells. BiFC analysis therefore provides a powerful approach for the visualization of a variety of processes that affect molecular proximity in living cells.
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影响因子：
46.9
作者：
Nagai, T;Ibata, K;Miyawaki, A
通讯作者：
Miyawaki, A
影响因子：
46.9
作者：
Galarneau, A;Primeau, M;Michnick, SW
通讯作者：
Michnick, SW
影响因子：
15
作者：
Magliery, TJ;Wilson, CGM;Regan, L
通讯作者：
Regan, L
影响因子：
5.3
作者：
Adolph, Doerte;Flach, Nadine;Ostareck-Lederer, Antje
通讯作者：
Ostareck-Lederer, Antje
影响因子：
3.9
作者：
Anderie, Ines;Schmid, Andreas
通讯作者：
Schmid, Andreas