TY - JOUR
T1 - Development of Fluorogenic Probes for Rapid High-Contrast Imaging of Transient Nuclear Localization of Sirtuin 3
AU - Gao, Jingchi
AU - Hori, Yuichiro
AU - Shimomura, Takashi
AU - Bordy, Mathieu
AU - Hasserodt, Jens
AU - Kikuchi, Kazuya
N1 - Funding Information:
This research was supported by JSPS KAKENHI (grant nos. JP17H06409 “Frontier Research on Chemical Communications”, JP18H03935, JP25220207 to K.K.; JP17H02210, JP18K19402, JP17H06005, JP18H04735 “Resonance Bio” to Y.H.; and JP19J12178 to J.G.), by JSPS Asian CORE Program, “Asian Chemical Biology Initiative”, by JSPS A3 Foresight Program, by the Japan Agency for Medical Research and Development (AMED, nos. 18he0902005h0004, 17ae0101041h9902, and 18fm0208018h0002 to K.K.), by JST SICORP (grant no. JPMJSC1602), by the Asahi Glass Foundation, by the Takeda Science Foundation, and by the French National Research Agency (ANR-16-JTIC-0001-01 to J.H.).
Publisher Copyright:
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
PY - 2020/3/2
Y1 - 2020/3/2
N2 - Protein labeling using fluorogenic probes enables the facile visualization of proteins of interest. Herein, we report new fluorogenic probes consisting of a rationally designed coumarin ligand for the live-cell fluorogenic labeling of the photoactive yellow protein (PYP)-tag. On the basis of the photochemical mechanisms of coumarin and the probe–tag interactions, we introduced a hydroxy group into an environment-sensitive coumarin ligand to modulate its spectroscopic properties and increase the labeling reaction rate. The resulting probe had a higher labeling reaction rate constant and a greater fluorescence OFF–ON ratio than any previously developed PYP-tag labeling probe. The probe enabled the fluorogenic labeling of intracellular proteins within minutes. Furthermore, we used our probe to investigate the localization of sirtuin 3 (SIRT3), a mitochondrial deacetylase. Although the nuclear localization of SIRT3 has been controversial, this transient nuclear localization was clearly captured by the rapid, high-contrast imaging enabled by our probe.
AB - Protein labeling using fluorogenic probes enables the facile visualization of proteins of interest. Herein, we report new fluorogenic probes consisting of a rationally designed coumarin ligand for the live-cell fluorogenic labeling of the photoactive yellow protein (PYP)-tag. On the basis of the photochemical mechanisms of coumarin and the probe–tag interactions, we introduced a hydroxy group into an environment-sensitive coumarin ligand to modulate its spectroscopic properties and increase the labeling reaction rate. The resulting probe had a higher labeling reaction rate constant and a greater fluorescence OFF–ON ratio than any previously developed PYP-tag labeling probe. The probe enabled the fluorogenic labeling of intracellular proteins within minutes. Furthermore, we used our probe to investigate the localization of sirtuin 3 (SIRT3), a mitochondrial deacetylase. Although the nuclear localization of SIRT3 has been controversial, this transient nuclear localization was clearly captured by the rapid, high-contrast imaging enabled by our probe.
UR - http://www.scopus.com/inward/record.url?scp=85074750751&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85074750751&partnerID=8YFLogxK
U2 - 10.1002/cbic.201900568
DO - 10.1002/cbic.201900568
M3 - Article
C2 - 31518474
AN - SCOPUS:85074750751
SN - 1439-4227
VL - 21
SP - 656
EP - 662
JO - ChemBioChem
JF - ChemBioChem
IS - 5
ER -