Unknown

Dataset Information

0

Design of a protein tag and fluorogenic probe with modular structure for live-cell imaging of intracellular proteins.


ABSTRACT: Conditional fluorescence imaging is a powerful technique for precise spatiotemporal analysis of proteins in live cells upon administration of a synthetic probe. To be applicable to various biological phenomena, probes must be membrane-permeable, have a high signal-to-noise ratio, and work quickly. To date, few probes meet all of these requirements. Here, we designed a fluorogenic probe (AcFCANB) that could label intracellular proteins fused to the photoactive yellow protein (PYP) tag in live cells within 30 min and used it to image heterochromatin protein 1 localization in nuclei. AcFCANB is based on a modular platform consisting of fluorophore, ligand and quencher. We accelerated the labeling reaction by strategic mutations of charged residues on the surface of PYP. A simple model based on molecular dynamics simulations quantitatively reproduced the cooperative effect of multiple mutations on labeling rate.

SUBMITTER: Kamikawa Y 

PROVIDER: S-EPMC5952543 | BioStudies | 2016-01-01

REPOSITORIES: biostudies

Similar Datasets

1000-01-01 | S-EPMC3213346 | BioStudies
2013-01-01 | S-EPMC3800097 | BioStudies
2016-01-01 | S-EPMC6273459 | BioStudies
2017-01-01 | S-EPMC5647265 | BioStudies
2020-01-01 | S-EPMC7175155 | BioStudies
2019-01-01 | S-EPMC6630177 | BioStudies
1000-01-01 | S-EPMC5452268 | BioStudies
2017-01-01 | S-EPMC5362743 | BioStudies
2019-01-01 | S-EPMC6662066 | BioStudies
2018-01-01 | S-EPMC5960302 | BioStudies