Development of fluorogenic probes for quick no-wash live-cell imaging of intracellular proteins.
We developed novel fluorogenic probes for no-wash live-cell imaging of proteins fused to PYP-tag, which is a small protein tag recently reported by our group. Through the design of a new PYP-tag ligand, specific intracellular protein labeling with rapid kinetics and fluorogenic response was accomplished. The probes crossed the cell membrane, and cytosolic and nuclear localizations of PYP-tagged proteins without cell washing were visualized within a 6-min reaction time. The fluorogenic response was due to the environmental effect of fluorophore upon binding to PYP-tag. Furthermore, the PYP-tag-based method was applied to the imaging of methyl-CpG-binding domain localization. This rapid protein-labeling system combined with the small protein tag and designed fluorogenic probes offers a powerful method to study the localization, movement, and function of cellular proteins.
Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan.
Yuichiro
Y
Hori
Tomoya
T
Norinobu
Motoki
M
Sato
Kyohei
K
Arita
Masahiro
M
Shirakawa
Kazuya
K
Kikuchi
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D002470:Cell Survival; D019175:DNA Methylation; D005456:Fluorescent Dyes / Q000737:chemistry*; D042541:Intracellular Space / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D041681:NIH 3T3 Cells; D020283:Photoreceptors, Microbial / Q000235:genetics; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D020283:Photoreceptors, Microbial; D011506:Proteins; C050162:photoactive yellow protein, Bacteria