BACKGROUND AND PURPOSEHydrogen sulfide (H 2 S) is a gasotransmitter produced from L-cysteine through the enzymatic action of cystathionine-γ-lyase (CSE) and/or cystathionine-β-synthase. D-Penicillamine is the D isomer of a dimethylated cysteine and has been used for the treatment of rheumatoid arthritis. As D-penicillamine is structurally very similar to cysteine, we have investigated whether D-penicillamine, as a cysteine analogue, has an effect on the H 2 S pathway.
EXPERIMENTAL APPROACHWe tested the effect of D-penicillamine (0.01-1 mM) in mouse aortic rings mounted in isolated organ baths and determined whether it could affect H 2 S biosynthesis. In particular, we investigated any possible inhibitor or donor behaviour by using recombinant enzyme-based assays and an in vivo approach.
KEY RESULTSD-Penicillamine, per se, showed little or no vasodilator effect, and it cannot be metabolized as a substrate in place of L-cysteine. However, D-penicillamine significantly reduced L-cysteine-induced vasodilatation in a concentration-dependent manner through inhibition of H 2 S biosynthesis, and this effect occurred at concentrations 10 times lower than those needed to induce the release of H 2 S. In particular, D-penicillamine selectively inhibited CSE in a pyridoxal-5′-phospate-dependent manner.
CONCLUSIONS AND IMPLICATIONSTaken together, our results suggest that D-penicillamine acts as a selective CSE inhibitor, leading to new perspectives in the design and use of specific pharmacological tools for H 2 S research. In addition, the inhibitory effect of D-penicillamine on CSE could account for its beneficial action in rheumatoid arthritis patients, where H 2 S has been shown to have a detrimental effect.
AbbreviationsMPST, 3-mercaptopyruvate sulfurtransferase; CBS, cystathionine-β-synthase; CSE, cystathionine-γ-lyase; DPD, N,N-dimethylp-phenylenediamine sulfate; D-pen, D-penicillamine; L-pen, L-penicillamine; H 2 S, hydrogen sulfide; IVM, intravital microscopy; PAG, D,L-propargylglycine; PE, phenylephrine; PLP, pyridoxal-5′-phosphate; RA, rheumatoid arthritis; TCA, trichloroacetic acid; ZnAc, zinc acetate
BJP
IntroductionHydrogen sulfide (H 2 S) is a gaseous molecule endogenously synthesized by cystathionine-γ-lyase (CSE), cystathionine-β-synthase (CBS) and 3-mercaptopyruvate sulfurtransferase (MPST). These enzymes are differently distributed throughout the human body and within cell compartments. MPST is mainly expressed in mitochondria (Stipanuk, 2004;Shibuya et al., 2009), while CSE and CBS show a wide distribution in diverse cell types. CSE and CBS can both metabolize the substrate L-cysteine to release H 2 S, and this reaction is strictly dependent upon the enzyme cofactor pyridoxal-5′-phosphate (PLP). The role of H 2 S has been widely investigated in several organs and tissues, and many studies have confirmed its crucial role in body physiology. For instance, CSE represents the prominent enzyme within the cardiovascular system, where H 2 S is a major player involved in regulating the function of heart and bloo...