title: Evidence for heme-mediated redox regulation of human cystathionine beta-synthase activity.
abstract: Human cystathionine beta-synthase catalyzes the first step in the catabolic removal of the toxic metabolite, homocysteine. It is unique in being dependent on both pyridoxal phosphate (PLP) and heme for activity. The reaction involves condensation of serine and homocysteine to give cystathionine. Although the role of PLP can be rationalized in analogy with other PLP-dependent enzymes that catalyze beta-replacement reactions, the role of the heme is unknown. In this study, we have purified and characterized the recombinant human enzyme and have examined the effect of heme oxidation state on enzyme activity. We find that under reducing conditions, generated by addition of titanium citrate, the enzyme exhibits a 1.7-fold lower activity than under oxidizing conditions. Reoxidation of the ferrous enzyme with ferricyanide results in alleviation of inhibition. This redox-linked change in enzyme activity correlates with changes in heme oxidation state monitored by UV-visible spectroscopy. Dithiothreitol, which does not reduce the enzyme-bound heme, does not perturb enzyme activity. These studies provide the first evidence for redox-linked regulation of cystathionine beta-synthase which is heme-dependent.
authors.affiliation: Department of Biochemistry, University of Nebraska, Lincoln, Nebraska 68588-0664, USA.
authors.forename: S
authors.initials: S
authors.lastname: Taoka
authors.forename: S
authors.initials: S
authors.lastname: Ohja
authors.forename: X
authors.initials: X
authors.lastname: Shan
authors.forename: W D
authors.initials: WD
authors.lastname: Kruger
authors.forename: R
authors.initials: R
authors.lastname: Banerjee
mesh_terms: D003541:Cystathionine beta-Synthase / Q000302:isolation & purification / Q000378:metabolism*; D006418:Heme / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D006418:Heme; D003541:Cystathionine beta-Synthase