Phosphonoacetaldehyde

Isbell, A. F.; Englert, Leo F.; Rosenberg, H.

doi:10.1021/jo01255a067

Cited by 40 publications

(20 citation statements)

References 2 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…PAld hydrolysis. This reaction was monitored at 25 °C by measuring the oxidation of β-NADH at 340 nm (ε = 6.22 mM −1 cm −1 ) in a 1 mL assay solutions initially containing PAc hydrolase, PAld (30), 10 u alcohol dehydrogenase, 0.5 mM β-NADH, 5 mM ZnCl 2 , and 5 mM MgCl 2 in 50 mM K + ADA (pH 7.0). Bis- p -nitrophenylphosphate, p -nitrophenylphosphate and p -nitrophenylsulfate hydrolysis .…”

Section: Methodsmentioning

confidence: 99%

Divergence of Chemical Function in the Alkaline Phosphatase Superfamily: Structure and Mechanism of the P−C Bond Cleaving Enzyme Phosphonoacetate Hydrolase

et al. 2011

View full text Add to dashboard Cite

Phosphonates constitute a class of natural products that mimic the properties of the more common organophosphate ester metabolite, yet are not readily degraded owing to the direct linkage of the phosphorus atom to the carbon atom. Phosphonate hydrolases have evolved to allow bacteria to utilize environmental phosphonates as a source of carbon and phosphorus. The work reported in this paper examines one such enzyme, phosphonoacetate hydrolase. By using a bioinformatic approach we circumscribed the biological range of phosphonoacetate hydrolase to a select group of bacterial species from different classes of Proteobacteria. In addition, using gene context we identified a novel 2-aminoethylphosphonate degradation pathway in which phosphonoacetate hydrolase is a participant. The X-ray structure of phosphonoformate-bound phosphonoacetate hydrolase was determined to reveal that this enzyme is most closely related to nucleotide pyrophosphatase/diesterase, a promiscuous two-zinc ion metalloenzyme of the alkaline phosphatase enzyme superfamily. The X-ray structure and metal ion specificity tests showed that phosphonoacetate hydrolase is also a two-zinc ion metalloenzyme. By using site-directed mutagenesis and 32P-labeling strategies, the catalytic nucleophile was shown to be Thr64. A structure-guided, site-directed mutation based inquiry of the catalytic contributions of active site residues identified Lys126 and Lys128 as the most likely candidates for stabilization of the aci-carboxylate dianion leaving group. A catalytic mechanism is proposed which combines Lys12/Lys128 leaving group stabilization with zinc ion activation of the Thr64 nucleophile and the substrate phosphoryl group.

show abstract

Section: Methodsmentioning

confidence: 99%

Divergence of Chemical Function in the Alkaline Phosphatase Superfamily: Structure and Mechanism of the P−C Bond Cleaving Enzyme Phosphonoacetate Hydrolase

et al. 2011

View full text Add to dashboard Cite

show abstract

“…The wildtype and mutant phosphonatase proteins were purified as previously described [10]. Pald was synthesized according to the published procedure [38]. Yeast alcohol dehydrogenase, NADH, and all buffers were purchased from Sigma.…”

Section: Methodsmentioning

confidence: 99%

Diversification of function in the haloacid dehalogenase enzyme superfamily: The role of the cap domain in hydrolytic phosphoruscarbon bond cleavage

Lahiri

Zhang

Dunaway‐Mariano

et al. 2006

Bioorganic Chemistry

View full text Add to dashboard Cite

Phosphonatase functions in the 2-aminoethylphosphonate (AEP) degradation pathway of bacteria, catalyzing the hydrolysis of the C-P bond in phosphonoacetaldehyde (Pald) via formation of a bicovalent Lys53ethylenamine/Asp12 aspartylphosphate intermediate. Because phosphonatase is a member of the haloacid dehalogenase superfamily, a family predominantly comprised of phosphatases, the question arises as to how this new catalytic activity evolved. The source of general acid-base catalysis for Schiff-base formation and aspartylphosphate hydrolysis was probed using pH-rate profile analysis of active-site mutants and X-ray crystallographic analysis of modified forms of the enzyme. The 2.9 Å X-ray crystal structure of the mutant Lys53Arg complexed with Mg +2 and phosphate shows that the equilibrium between the open and the closed conformation is disrupted, favoring the open conformation. Thus, proton dissociation from the cap domain Lys53 is required for cap domain-core domain closure. The likely recipient of the Lys53 proton is a water-His56 pair that serves to relay the proton to the carbonyl oxygen of the phosphonoacetaldehyde (Pald) substrate upon addition of the Lys53. The pH-rate profile analysis of active-site mutants was carried out to test this proposal. The proximal core domain residues Cys22 and Tyr128 were ruled out, and the role of cap domain His56 was supported by the results. The X-ray crystallographic structure of wild-type phosphonatase reduced with NaBH 4 in the presence of Pald was determined at 2.4 Å resolution to reveal Nε-ethyl-Lys53 juxtaposed with a sulfate ligand bound in the phosphate site. The position of the C(2) of the N-ethyl group in this structure is consistent with the hypothesis that the cap domain Nε-ethylenamine-Lys53 functions as a general base in the hydrolysis of the aspartylphosphate bicovalent enzyme intermediate. Because the enzyme residues proposed to play a key role in P-C bond cleavage are localized on the cap domain, this domain appears to have evolved to support the diversification of the HAD phosphatase core domain for catalysis of hydrolytic P-C bond cleavage.

show abstract

“…Chemical Synthesis of Substrates-PnAA, HEP, and hydroxymethylphosphonate (HMP) were synthesized according to the methods reported elsewhere (14,19).…”

Section: Methodsmentioning

confidence: 99%

Biosynthesis of 2-Hydroxyethylphosphonate, an Unexpected Intermediate Common to Multiple Phosphonate Biosynthetic Pathways

Shao

Blodgett

Circello

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

Phosphonic acids encompass a common yet chemically diverse class of natural products that often possess potent biological activities. Here we report that, despite the significant structural differences among many of these compounds, their biosynthetic routes contain an unexpected common intermediate, 2-hydroxyethyl-phosphonate, which is synthesized from phosphonoacetaldehyde by a distinct family of metal-dependent alcohol dehydrogenases (ADHs). Although the sequence identity of the ADH family members is relatively low (34 -37%), in vitro biochemical characterization of the homologs involved in biosynthesis of the antibiotics fosfomycin, phosphinothricin tripeptide, and dehydrophos (formerly A53868) unequivocally confirms their enzymatic activities. These unique ADHs have exquisite substrate specificity, unusual metal requirements, and an unprecedented monomeric quaternary structure. Further, sequence analysis shows that these ADHs form a monophyletic group along with additional family members encoded by putative phosphonate biosynthetic gene clusters. Thus, the reduction of phosphonoacetaldehyde to hydroxyethyl-phosphonate may represent a common step in the biosynthesis of many phosphonate natural products, a finding that lends insight into the evolution of phosphonate biosynthetic pathways and the chemical structures of new C-P containing secondary metabolites.

show abstract

Phosphonoacetaldehyde

Cited by 40 publications

References 2 publications

Divergence of Chemical Function in the Alkaline Phosphatase Superfamily: Structure and Mechanism of the P−C Bond Cleaving Enzyme Phosphonoacetate Hydrolase

Divergence of Chemical Function in the Alkaline Phosphatase Superfamily: Structure and Mechanism of the P−C Bond Cleaving Enzyme Phosphonoacetate Hydrolase

Diversification of function in the haloacid dehalogenase enzyme superfamily: The role of the cap domain in hydrolytic phosphoruscarbon bond cleavage

Biosynthesis of 2-Hydroxyethylphosphonate, an Unexpected Intermediate Common to Multiple Phosphonate Biosynthetic Pathways

Contact Info

Product

Resources

About