Split mCherry as a new red bimolecular fluorescence complementation system for visualizing protein–protein interactions in living cells

• Published in: Biochemical and biophysical research communications Vol. 367; no. 1; pp. 47 - 53
• Main Authors: Fan, Jin-Yu, Cui, Zong-Qiang, Wei, Hong-Ping, Zhang, Zhi-Ping, Zhou, Ya-Feng, Wang, Yun-Peng, Zhang, Xian-En
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 29.02.2008
• Subjects: Amino Acid Sequence; Animals; Antigens, Polyomavirus Transforming - chemistry; Antigens, Polyomavirus Transforming - metabolism; Base Sequence; BiFC; Bimolecular fluorescence complementation; Biosensing Techniques - methods; Cells, Cultured - metabolism; Cells, Cultured - pathology; Chlorocebus aethiops; Fluorescence Resonance Energy Transfer - methods; Fluorescent Dyes - chemistry; Humans; Leukemia, Promyelocytic, Acute - metabolism; Leukemia, Promyelocytic, Acute - pathology; Luminescent Proteins - chemistry; mCherry; Molecular Sequence Data; Protein–protein interactions; Red Fluorescent Protein; Simian virus 40; Tumor Suppressor Protein p53 - chemistry; Tumor Suppressor Protein p53 - metabolism; Vero Cells - metabolism; Vero Cells - pathology; mCherry; Protein–protein interactions; BiFC; Bimolecular fluorescence complementation; Red fluorescent protein
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2007.12.101

Bimolecular fluorescence complementation (BiFC) is a recently developed technique for detection of protein–protein interactions in living cells. In this study, a new red BiFC system was developed by splitting mCherry, a mutant monomeric red fluorescent protein, into two fragments between amino acids 159–160 and was verified using a pair of interacting proteins, SV40 large T antigen (LTag), and human p53 protein. By combined use of the mCherry-based red BiFC system with a Venus-based yellow BiFC system, the interaction between LTag and p53 as well as the interaction between sp100 and promyelocytic leukemia protein (PML), were detected simultaneously in Vero cells. The brilliant redness, short maturation time, and the long excitation and emission wavelengths (587/610 nm) of mCherry make the new BiFC system an excellent candidate for analyzing protein–protein interactions in living cells and for studying multiple protein–protein interactions when coupled with other BiFC systems.