1 research outputs found
Oxyluciferin Derivatives: A Toolbox of Environment-Sensitive Fluorescence Probes for Molecular and Cellular Applications
In this work, we
used firefly oxyluciferin (<b>OxyLH</b><sub><b>2</b></sub>) and its polarity-dependent fluorescence mechanism
as a sensitive tool to monitor biomolecular interactions. The chromophores, <b>OxyLH</b><sub><b>2</b></sub>, and its two analogues, <b>4-MeOxyLH</b> and <b>4,6′-DMeOxyL</b>, were modified
trough carboxylic functionalization and then coupled to the N-terminus
part of Tat and NCp7 peptides of human immunodeficiency virus type-1
(HIV-1). The photophysical properties of the labeled peptides were
studied in live cells as well as in complex with different oligonucleotides
in solution. By monitoring the emission properties of these derivatives
we were able, for the first time, to study <i>in vitro</i> biomolecular interactions using oxyluciferin as a sensor. As an
additional application, cyclopropyl-oxyluciferin (<b>5,5-Cpr-OxyLH</b>) was site-specifically conjugated to the thiol group (Cys-232) of
the human protein α-1 antytripsin to investigate its interaction
with porcine pancreatic elastase. Our data demonstrate that <b>OxyLH</b><sub><b>2</b></sub> and its derivatives can be
used as fluorescence reporters for monitoring biomolecular interactions