Phosphonomycin: Structure and Synthesis

Christensen, B. G.; Leanza, W. J.; Beattie, Thomas R.; Patchett, A. A.; Arison, Byron H.; Ormond, Robert E.; Kuehl, Frederick A.; Albers-Schönberg, Georg; Jardetzky, Oleg

doi:10.1126/science.166.3901.123

Cited by 176 publications

(97 citation statements)

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“…1,2 It irreversibly inhibits MurA (UDP-N-acetylglucosamine-3-enolpyruvyltransferase), an important enzyme in bacterial cell wall biosynthesis (Scheme 1). 3 This inhibition is achieved through alkylation of an active-site cysteine residue, which performs a nucleophilic attack on the C2 atom of fosfomycin.…”

Section: Introductionmentioning

confidence: 99%

Determinants of Regioselectivity and Chemoselectivity in Fosfomycin Resistance Protein FosA from QM/MM Calculations

Liao

Thiel

2013

J. Phys. Chem. B

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FosA is a manganese-dependent enzyme that utilizes a Mn 2+ ion to catalyze the inactivation of the fosfomycin antibiotic by glutathione (GSH) addition. We report a theoretical study on the catalytic mechanism and the factors governing the regioselectivity and chemoselectivity of FosA. Density functional theory (DFT) calculations on the uncatalyzed reaction give high barriers and almost no regioselectivity even when adding two water molecules to assist the proton transfer. According to quantum mechanics/molecular mechanics (QM/MM) calculations on the full solvated protein, the enzyme-catalyzed glutathione addition reaction involves two major chemical steps that both proceed in the sextet state: proton transfer from the GSH thiol group to the Tyr39 anion and nucleophilic attack by the GSH thiolate leading to epoxide ring-opening. The second step is rate-limiting and is facilitated by the presence of the high-spin Mn 2+ ion that functions as a Lewis acid and stabilizes the leaving oxyanion through direct coordination. The barrier for C1 attack is computed to be 8.9 kcal/mol lower than that for C2 attack, in agreement with the experimentally observed regioselectivity of the enzyme. Further QM/MM calculations on the alternative water attack predict a concerted mechanism for this reaction, where the deprotonation of water, nucleophilic attack, and epoxide ring-opening take place via the same transition state. The calculated barrier is 8.3 kcal/mol higher than that for GSH attack, in line with the observed chemoselectivity of the enzyme, which manages to catalyze the addition of GSH in the presence of water molecules around its active site. The catalytic efficiency, regioselectivity, and chemoselectivity of FosA are rationalized in terms of the influence of the active-site protein environment and the different stabilization of the distorted substrates in the relevant transition states.

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Section: Introductionmentioning

confidence: 99%

Determinants of Regioselectivity and Chemoselectivity in Fosfomycin Resistance Protein FosA from QM/MM Calculations

Liao

Thiel

2013

J. Phys. Chem. B

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“…Fosfomycin is a broad-spectrum antibiotic produced by some strains of Streptomyces (Christensen et al, 1969 ;Hendlin et al, 1969). Its mechanism of action was first described by Kahan et al (1974), who showed that it inhibits the enzyme UDP-N-acetylglucosamine enolpyruvyl transferase (MurA), which transfers the enolpyruvyl moiety of phosphoenolpyruvate to the 3h hydroxyl group of UDP-N-acetylglucosamine, the first committed step in bacterial peptidoglycan biosynthesis.…”

Section: Introductionmentioning

confidence: 99%

Alteration of a single amino acid residue reverses fosfomycin resistance of recombinant MurA from Mycobacterium tuberculosis The EMBL accession number for the sequence in this paper is X96711.

Smet¹,

Kempsell²,

Gallagher³

et al. 1999

Microbiology

131

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Mycobacterium tuberculosis has innate resistance to a range of broadspectrum antimicrobial agents. This may in part reflect the relative impermeability of the mycobacterial cell wall, but additional specific mechanisms may also be important. In the case of fosfomycin, it has been suggested that a key difference in the active site of the M. tuberculosis MurA enzyme might confer resistance. In Escherichia coli, fosfomycin covalently binds to a cysteine normally involved in the enzymic activity, while protein alignments predict an aspartate at this position in the M. tuberculosis MurA. In the present study, it is demonstrated that the wild-type M. tuberculosis MurA is indeed resistant to fosfomycin, and that it becomes sensitive following replacement of the aspartate residue in position 117 by a cysteine. In addition, the study illustrates the use of an inducible expression system in mycobacteria to allow functional characterization of an M. tuberculosis enzyme that is unstable during constitutive expression.

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“…1). 8,9) Its target is known as the enzyme forming UDP-Nacetylmuramic acid from UDP-N-acetyl glucosamine and phosphoenolpyruvic acid. 10) Thus fosfomycin is an antibiotic that inhibits bacterial peptidoglycan synthesis.…”

mentioning

confidence: 99%

Fosmidomycin Resistance in Adenylate Cyclase Deficient (cya) Mutants ofEscherichia coli

Sakamoto

Furukawa

Ogihara

et al. 2003

Bioscience, Biotechnology, and Biochemistry

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Phosphonomycin: Structure and Synthesis

Abstract: Synthesis and resolution of the antibiotic phosphonomycin are described. The structure is (-)(IR, 2S)-1,2-epoxypropylphosphonic acid.

Cited by 176 publications

References 5 publications

Determinants of Regioselectivity and Chemoselectivity in Fosfomycin Resistance Protein FosA from QM/MM Calculations

Determinants of Regioselectivity and Chemoselectivity in Fosfomycin Resistance Protein FosA from QM/MM Calculations

Alteration of a single amino acid residue reverses fosfomycin resistance of recombinant MurA from Mycobacterium tuberculosis The EMBL accession number for the sequence in this paper is X96711.

Fosmidomycin Resistance in Adenylate Cyclase Deficient (cya) Mutants ofEscherichia coli

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