Evaluation of a dithiocarbamate derivative as an inhibitor of human glutaredoxin-1.
BACKGROUND
Glutaredoxins (GRX) are involved in the regulation of thiol redox state. GRX-1 is a cytosolic enzyme responsible for the catalysis of deglutathionylation of proteins. To date, very few inhibitors of GRX-1 have been reported.


OBJECTIVE
The objective of this paper is to report 2-acetylamino-3-[4-(2-acetylamino-2-carboxyethyl-sulfanylthiocarbonylamino)phenylthiocarbamoylsulfanyl]propionic acid (2-AAPA) as an inhibitor of human GRX-1.


METHODS
The mechanism of inhibition of GRX-1 was investigated using dialysis, substrate protection, and mass spectrometry.


RESULTS
2-AAPA inhibits GRX-1 in a time and concentration dependent manner. The activity did not return following dialysis indicating that inhibition is irreversible. Results of substrate protection and mass spectrometry indicate that the inhibition is occurring at the active site. The compound also produced GRX inhibition in human ovarian cancer cells.


CONCLUSIONS
2-AAPA is an irreversible GRX-1 inhibitor with similar or greater potency compared to previously reported inhibitors.


CONCLUSIONS
The inhibition of GRX-1 by 2-AAPA could be used as a tool to study thiol redox state.
Department of Pharmaceutical Sciences, College of Pharmacy, South Dakota State University, Brookings, SD, USA.
Satya S
SS
Sadhu
Eduardo
E
Callegari
Yong
Y
Zhao
Xiangming
X
Guan
Teresa
T
Seefeldt
D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D045744:Cell Line, Tumor / Q000187:drug effects; D003956:Dialysis; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D054477:Glutaredoxins / Q000037:antagonists & inhibitors*; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D013859:Thiocarbamates / Q000737:chemistry / Q000494:pharmacology*
C539612:2-acetylamino-3-(4-(2-acetylamino-2-carboxyethylsulfanylthiocarbonylamino)phenylthiocarbamoylsulfanyl)propionic acid; D004791:Enzyme Inhibitors; C516005:GLRX protein, human; D054477:Glutaredoxins; D013859:Thiocarbamates; D005978:Glutathione; D000111:Acetylcysteine