5 research outputs found
Media 1: Independent and simultaneous three-dimensional optical trapping and imaging
Originally published in Biomedical Optics Express on 01 October 2013 (boe-4-10-2087
Media 3: Tomographic phase microscopy with 180° rotation of live cells in suspension by holographic optical tweezers
Originally published in Optics Letters on 15 April 2015 (ol-40-8-1881
Media 4: Tomographic phase microscopy with 180° rotation of live cells in suspension by holographic optical tweezers
Originally published in Optics Letters on 15 April 2015 (ol-40-8-1881
Real-Time Imaging of the Azole Class of Antifungal Drugs in Live Candida Cells
Azoles are the most
commonly used class of antifungal drugs, yet
where they localize within fungal cells and how they are imported
remain poorly understood. Azole antifungals target lanosterol 14α-demethylase,
a cytochrome P450, encoded by <i>ERG11</i> in <i>Candida
albicans</i>, the most prevalent fungal pathogen. We report the
synthesis of fluorescent probes that permit visualization of antifungal
azoles within live cells. Probe <b>1</b> is a dansyl dye-conjugated
azole, and probe <b>2</b> is a Cy5-conjugated azole. Docking
computations indicated that each of the probes can occupy the active
site of the target cytochrome P450. Like the azole drug fluconazole,
probe <b>1</b> is not effective against a mutant that lacks
the target cytochrome P450. In contrast, the azole drug ketoconazole
and probe <b>2</b> retained some antifungal activity against
mutants lacking the target cytochrome P450, implying that both act
against more than one target. Both fluorescent azole probes colocalized
with the mitochondria, as determined by fluorescence microscopy with
MitoTracker dye. Thus, these fluorescent probes are useful molecular
tools that can lead to detailed information about the activity and
localization of the important azole class of antifungal drugs