10.1021/acs.analchem.9b04552.s001
Yahya Alqahtani
Yahya
Alqahtani
Shenggang Wang
Shenggang
Wang
Yue Huang
Yue
Huang
Asim Najmi
Asim
Najmi
Xiangming Guan
Xiangming
Guan
Design, Synthesis, and Characterization of Bis(7‑(<i>N</i>‑(2-morpholinoethyl)sulfamoyl)benzo[<i>c</i>][1,2,5]oxadiazol-5-yl)sulfane for Nonprotein Thiol Imaging in Lysosomes
in Live Cells
American Chemical Society
2019
subcellular thiol density
LysoTracker Red DND -99
cell nonprotein thiol
OH
COOH
lysosomal thiols
NPSH
subcellular organelle lysosome
BISMORX
NH
Nonprotein Thiol Imaging
Live Cells Thiols
lysosomal degradation process
2019-11-08 22:43:55
Journal contribution
https://acs.figshare.com/articles/journal_contribution/Design_Synthesis_and_Characterization_of_Bis_7_i_N_i_2-morpholinoethyl_sulfamoyl_benzo_i_c_i_1_2_5_oxadiazol-5-yl_sulfane_for_Nonprotein_Thiol_Imaging_in_Lysosomes_in_Live_Cells/10278050
Thiols
are critical to cellular structures and functions. Disturbance
of cellular thiols has been found to affect cell functions and cause
various diseases. Intracellularly, thiols were found unevenly distributed
in subcellular organelles. Probes capable of detecting subcellular
thiol density in live cells are valuable tools in determining thiols’
roles at the subcellular level. The subcellular organelle lysosome
is the place where unwanted macromolecules are removed through degradation
by hydrolytic enzymes. The degradation also serves as a regulation
of a variety of cellular functions such as autophagy, endocytosis,
and phagocytosis to maintain cellular homeostasis. Thiols are found
to be involved in the lysosomal degradation process. A probe that
can detect lysosomal thiols in live cells will be a valuable tool
in unveiling the roles of thiols in lysosomes. We would like to report
bis(7-(<i>N</i>-(2-morpholinoethyl)sulfamoyl)benzo[<i>c</i>][1,2,5]-oxadiazol-5-yl)sulfane (BISMORX) as a thiol specific
fluorogenic agent for live cell nonprotein thiol (NPSH) imaging in
lysosomes through fluorescence microscopy. BISMORX itself shows no
fluorescence and reacts readily with a NPSH to form a fluorescent
thiol adduct with excitation and emission wavelengths of 380 and 540
nm, respectively. BISMORX does not react with compounds containing
nucleophilic functional groups other than thiols such as −OH,
−NH<sub>2</sub>, and −COOH. No reaction was observed
either when BISMORX was mixed with protein thiols. BISMORX was able
to image, quantify, and detect the change of NPSH in lysosomes in
live cells. A colocalization experiment with LysoTracker Red DND-99
confirmed that the thiols imaged by BISMORX were indeed lysosomal
thiols.