A novel far-red bimolecular fluorescence complementation system that allows for efficient visualization of protein interactions under physiological conditions

作者：Chu, Jun; Zhang, Zhihong; Zheng, Ying; Yang, Jie; Qin, Lingsong; Lu, Jinling; Huang, Zhen-Li; Zeng, Shaoqun; Luo, Qingming^*

来源：Biosensors and Bioelectronics, 2009, 25(1): 234-239.

DOI：10.1016/j.bios.2009.06.008

• Protein-protein interactions
• Bimolecular fluorescence complementation (BiFC)
• Far-red fluorescent protein
• mKate
• Multicolor

Fluorescent protein (FP) has enabled the analysis of biomolecular interactions in living cells,and bimolecular fluorescence complementation (BiFC) represents one of the newly developed imaging technologies to directly visualize protein-protein interactions in living cells. Although 10 different FPs that cover a broad range of spectra have been demonstrated to support BiFC, only Cerulean (cyan FP variant), Citrine and Venus (yellow FP variants)-based BiFC systems can be used under 37 degrees C physiological temperature. The sensitivity of two mRFP-based red BiFC systems to higher temperatures (i.e., 37 degrees C) limits their applications in most mammalian cell-based studies. Here we report that mLumin, a newly isolated far-red fluorescent protein variant of mKate with an emission maximum of 621 nm, enables BiK analysis of protein-protein interactions at 37 degrees C in living mammalian cells. Furthermore, the combination of mLumin with Cerulean- and Venus-based BiFC systems allows for simultaneous visualization of three pairs of protein-protein interactions in the same cell. The mLumin-based BiK system will facilitate simultaneous visualization of multiple protein-protein interactions in living cells and offer the potential to visualize protein-protein interactions in living animals.

• 出版日期2009-9-15
• 单位华中科技大学