Although many protein
labeling probes have been developed to elucidate
the trafficking and turnover processes of cell surface proteins, real-time
tracking of intracellular proteins remains a challenging task. Herein,
we describe a new design to construct a cell-permeable, photostable,
and far-red fluorescent turn-on probe to enable no-wash, organelle-specific,
and long-term visualization of intracellular SNAP-tagged proteins
in living cells. When the probe was used in dual-color pulse chase
labeling experiments to differentiate between preLamin and mature
Lamin, our results reveal that the shape of mature Lamin can be altered
by the newly synthesized preLamin and that this alteration is progressive,
cumulative, and due to a concentration-dependent dominant-negative
effect of preLamin. We believe that this probe can also be applied
to other intracellular proteins whose cellular localization and synthesis
changes dynamically in response to external stimuli.
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