Isozyme-specific fluorescent inhibitor of glutathione s-transferase omega 1.
Recently, the glutathione S-transferase omega 1 (GSTO1) is suspected to be involved in certain cancers and neurodegenerative diseases. However, profound investigation on the pathological roles of GSTO1 has been hampered by the lack of specific methods to determine or modulate its activity in biological systems containing other isoforms with similar catalytic function. Here, we report a fluorescent compound that is able to inhibit and monitor the activity of GSTO1. We screened 43 fluorescent chemicals and found a compound (6) that binds specifically to the active site of GSTO1. We observed that compound 6 inhibits GSTO1 by covalent modification but spares other isoforms in HEK293 cells and demonstrated that compound 6 could report the activity of GSTO1 in NIH/3T3 or HEK293 cells by measuring the fluorescence intensity of the labeled amount of GSTO1 in SDS-PAGE. Compound 6 is a useful tool to study GSTO1, applicable as a specific inhibitor and an activity reporter.
Son
Junghyun
J
Lee
Jae-Jung
JJ
Lee
Jun-Seok
JS
Schüller
Andreas
A
Chang
Young-Tae
YT
D000818:Animals; D020134:Catalytic Domain; D002460:Cell Line; D002850:Chromatography, Gel; D056148:Chromatography, Reverse-Phase; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes; D005982:Glutathione Transferase / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D007527:Isoenzymes / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D041681:NIH 3T3 Cells; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D007527:Isoenzymes; C490476:GSTO1 protein, human; D005982:Glutathione Transferase