Journal
BIOSENSORS & BIOELECTRONICS
Volume 25, Issue 1, Pages 234-239Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2009.06.008
Keywords
Protein-protein interactions; Bimolecular fluorescence complementation (BiFC); Far-red fluorescent protein; mKate; Multicolor
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Funding
- National Natural Science Foundation of China [30770525, 30727002]
- National High-Tech R&D Program of China [2006AA020801]
- Program for Changjiang Scholars and Innovative Research Team in University
- New Century Excellent Talents in University
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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. (C) 2009 Elsevier B.V. All rights reserved.
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