title: Comparative study of enzyme activity and heme reactivity in Drosophila melanogaster and Homo sapiens cystathionine β-synthases.
abstract: Cystathionine β-synthase (CBS) is the first and rate-limiting enzyme in the transsulfuration pathway, which is critical for the synthesis of cysteine from methionine in eukaryotes. CBS uses coenzyme pyridoxal 5'-phosphate (PLP) for catalysis, and S-adenosylmethionine regulates the activity of human CBS, but not yeast CBS. Human and fruit fly CBS contain heme; however, the role for heme is not clear. This paper reports biochemical and spectroscopic characterization of CBS from fruit fly Drosophila melanogaster (DmCBS) and the CO/NO gas binding reactions of DmCBS and human CBS. Like CBS enzymes from lower organisms (e.g., yeast), DmCBS is intrinsically highly active and is not regulated by AdoMet. The DmCBS heme coordination environment, the reactivity, and the accompanying effects on enzyme activity are similar to those of human CBS. The DmCBS heme bears histidine and cysteine axial ligands, and the enzyme becomes inactive when the cysteine ligand is replaced. The Fe(II) heme in DmCBS is less stable than that in human CBS, undergoing more facile reoxidation and ligand exchange. In both CBS proteins, the overall stability of the protein is correlated with the heme oxidation state. Human and DmCBS Fe(II) hemes react relatively slowly with CO and NO, and the rate of the CO binding reaction is faster at low pH than at high pH. Together, the results suggest that heme incorporation and AdoMet regulation in CBS are not correlated, possibly providing two independent means for regulating the enzyme.
authors.affiliation: Department of Chemistry, University of Wisconsin-Madison, 1101 University Avenue, Madison, WI 53706, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Su
authors.forename: Tomas
authors.initials: T
authors.lastname: Majtan
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Freeman
authors.forename: Rachel
authors.initials: R
authors.lastname: Linck
authors.forename: Sarah
authors.initials: S
authors.lastname: Ponter
authors.forename: Jan P
authors.initials: JP
authors.lastname: Kraus
authors.forename: Judith N
authors.initials: JN
authors.lastname: Burstyn
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002248:Carbon Monoxide / Q000737:chemistry; D017124:Conserved Sequence; D003541:Cystathionine beta-Synthase / Q000737:chemistry* / Q000235:genetics; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics; D004331:Drosophila melanogaster / Q000201:enzymology*; D004795:Enzyme Stability; D019143:Evolution, Molecular; D006418:Heme / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000737:chemistry; D010084:Oxidation-Reduction; D011485:Protein Binding
chemical_list: D029721:Drosophila Proteins; D009569:Nitric Oxide; D006418:Heme; D002248:Carbon Monoxide; D003541:Cystathionine beta-Synthase