title: Analogs of 1-phosphonooxy-2,2-dihydroxy-3-oxo-5-(methylthio)pentane, an acyclic intermediate in the methionine salvage pathway: a new preparation and characterization of activity with E1 enolase/phosphatase from Klebsiella oxytoca.
abstract: The methionine salvage pathway allows the in vivo recovery of the methylthio moiety of methionine upon the formation of methylthioadenosine (MTA) from S-adenosylmethionine (SAM). The Fe(II)-containing form of acireductone dioxygenase (ARD) catalyzes the penultimate step in the pathway in Klebsiella oxytoca, the oxidative cleavage of the acireductone 1,2-dihydroxy-3-oxo-5-(methylthio)pent-1-ene (2) by dioxygen to give formate and 2-oxo-4-(methylthio)butyrate (3). The Ni(II)-bound form (Ni-ARD) catalyzes an off-pathway shunt, forming 3-(methylthio)propionate (4), carbon monoxide, and formate. Acireductone 2 is formed by the action of another enzyme, E1 enolase/phosphatase, on precursor 1-phosphonooxy-2,2-dihydroxy-3-oxo-5-methylthiopentane (1). Simple syntheses of several analogs of 1 are described, and their activity as substrates for E1 enolase/phosphatase characterized. A new bacterial overexpression system and purification procedure for E1, a member of the haloacid dehalogenase (HAD) superfamily, is described, and further characterization of the enzyme presented.
authors.affiliation: Department of Chemistry, Brandeis University, 415 South Street, MS 015, Waltham, MA 02454-9110, USA.
authors.forename: Yalin
authors.initials: Y
authors.lastname: Zhang
authors.forename: Melissa H
authors.initials: MH
authors.lastname: Heinsen
authors.forename: Milka
authors.initials: M
authors.lastname: Kostic
authors.forename: Gina M
authors.initials: GM
authors.lastname: Pagani
authors.forename: Thomas V
authors.initials: TV
authors.lastname: Riera
authors.forename: Iva
authors.initials: I
authors.lastname: Perovic
authors.forename: Lizbeth
authors.initials: L
authors.lastname: Hedstrom
authors.forename: Barry B
authors.initials: BB
authors.lastname: Snider
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Pochapsky
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D017931:DNA Primers; D041121:Klebsiella oxytoca / Q000201:enzymology*; D009682:Magnetic Resonance Spectroscopy; D008715:Methionine / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010755:Organophosphates / Q000737:chemistry*; D010420:Pentanes / Q000737:chemistry*; D010751:Phosphopyruvate Hydratase / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013056:Spectrophotometry, Ultraviolet
chemical_list: C498106:1-phosphonooxy-2,2-dihydroxy-3-oxo-5-(methylthio)pentane; D017931:DNA Primers; D010755:Organophosphates; D010420:Pentanes; D008715:Methionine; D010744:Phosphoric Monoester Hydrolases; D010751:Phosphopyruvate Hydratase