Split mCherry as a new red bimolecular fluorescence complementation system for visualizing protein-protein interactions in living cells.
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.
Fan
Jin-Yu
JY
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, 44#, Xiao Hong Shan, Wuhan 430071, China.
Cui
Zong-Qiang
ZQ
Wei
Hong-Ping
HP
Zhang
Zhi-Ping
ZP
Zhou
Ya-Feng
YF
Wang
Yun-Peng
YP
Zhang
Xian-En
XE
D000595:Amino Acid Sequence; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D002478:Cells, Cultured / Q000378:metabolism / Q000473:pathology; D002522:Chlorocebus aethiops; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism / Q000473:pathology; D008164:Luminescent Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000378:metabolism; D014709:Vero Cells / Q000378:metabolism / Q000473:pathology; D000097573:Red Fluorescent Protein
D000952:Antigens, Polyomavirus Transforming; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D016159:Tumor Suppressor Protein p53; D000097573:Red Fluorescent Protein