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Selectivity of commonly used pharmacological inhibitors for cystathionine β synthase ( CBS ) and cystathionine γ lyase ( CSE )
Author(s) -
Asimakopoulou Antonia,
Panopoulos Panagiotis,
Chasapis Christos T,
Coletta Ciro,
Zhou Zongmin,
Cirino Giuseppe,
Giannis Athanassios,
Szabo Csaba,
Spyroulias Georgios A,
Papapetropoulos Andreas
Publication year - 2013
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1111/bph.12171
Subject(s) - cystathionine beta synthase , aminooxyacetic acid , cystathionine gamma lyase , hydroxylamine , chemistry , cysteine , enzyme , biochemistry , fusion protein , microbiology and biotechnology , recombinant dna , biology , gene
Background and Purpose Hydrogen sulfide ( H 2 S ) is a signalling molecule that belongs to the gasotransmitter family. Two major sources for endogenous enzymatic production of H 2 S are cystathionine β synthase ( CBS ) and cystathionine γ lyase ( CSE ). In the present study, we examined the selectivity of commonly used pharmacological inhibitors of H 2 S biosynthesis towards CSE and CBS . Experimental Approach To address this question, human CSE or CBS enzymes were expressed and purified from E scherichia coli as fusion proteins with GSH‐ S ‐transferase. After purification, the activity of the recombinant enzymes was tested using the methylene blue method. Key Results β‐cyanoalanine ( BCA ) was more potent in inhibiting CSE than propargylglycine ( PAG ) ( IC 50 14 ± 0.2 μM vs. 40 ± 8 μM respectively). Similar to PAG , L ‐aminoethoxyvinylglycine ( AVG ) only inhibited CSE , but did so at much lower concentrations. On the other hand, aminooxyacetic acid ( AOAA ), a frequently used CBS inhibitor, was more potent in inhibiting CSE compared with BCA and PAG ( IC 50 1.1 ± 0.1 μM); the IC 50 for AOAA for inhibiting CBS was 8.5 ± 0.7 μM. In line with our biochemical observations, relaxation to L‐cysteine was blocked by AOAA in aortic rings that lacked CBS expression. Trifluoroalanine and hydroxylamine, two compounds that have also been used to block H 2 S biosynthesis, blocked the activity of CBS and CSE . Trifluoroalanine had a fourfold lower IC 50 for CBS versus CSE , while hydroxylamine was 60‐fold more selective against CSE . Conclusions and Implications In conclusion, although PAG , AVG and BCA exhibit selectivity in inhibiting CSE versus CBS , no selective pharmacological CBS inhibitor is currently available.