Production of the neuromodulator H2S by cystathionine beta-synthase via the condensation of cysteine and homocysteine.
Hydrogen sulfide (H2S) has been observed in relatively high concentrations in the mammalian brain and has been shown to act as a neuromodulator. However, there is confusion in the literature regarding the actual source of H2S production. Reactions catalyzed by the cystathionine beta-synthase enzyme (CBS) are one possible source for the production of H2S. Here we show that the CBS enzyme can efficiently produce H2S via a beta-replacement reaction in which cysteine is condensed with homocysteine to form cystathionine and H2S. The production of H2S by this reaction is at least 50 times more efficient than that produced by hydrolysis of cysteine alone via beta-elimination. Kinetic studies demonstrate that the Km and Kcat for cysteine is 3-fold higher and 2-fold lower, respectively, than that for serine. Consistent with these data, in vitro reconstitution studies show that at physiologically relevant concentrations of serine, homocysteine, and cysteine, about 5% of the cystathionine formed is from cysteine. We also show that AdoMet stimulates this H2S producing reaction but that there is no evidence for stimulation by calcium and calmodulin as reported previously. In summary, these results confirm the ability of CBS to produce H2S, but show in contrast to prior reports that the major mechanism is via beta-replacement and not cysteine hydrolysis. In addition, these studies provide a biochemical explanation for the previously inexplicable homocysteine-lowering effects of N-acetylcysteine treatments in humans.
Chen
Xulin
X
Division of Population Science, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA.
Jhee
Kwang-Hwan
KH
Kruger
Warren D
WD
D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000494:pharmacology; D003541:Cystathionine beta-Synthase / Q000235:genetics / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006710:Homocysteine / Q000378:metabolism; D006801:Humans; D006862:Hydrogen Sulfide / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D018377:Neurotransmitter Agents / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
D002147:Calmodulin; D018377:Neurotransmitter Agents; D011994:Recombinant Proteins; D006710:Homocysteine; D003541:Cystathionine beta-Synthase; D003545:Cysteine; D002118:Calcium; D006862:Hydrogen Sulfide