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.
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.
Zhang
Yalin
Y
Department of Chemistry, Brandeis University, 415 South Street, MS 015, Waltham, MA 02454-9110, USA.
Heinsen
Melissa H
MH
Kostic
Milka
M
Pagani
Gina M
GM
Riera
Thomas V
TV
Perovic
Iva
I
Hedstrom
Lizbeth
L
Snider
Barry B
BB
Pochapsky
Thomas C
TC
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
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