Fluorine as a hydroxy analogue. Stereospecific phosphorylation of 2-deosy-2-fluoroglycerol by glucerol kinase

Briley, Patricia A.; Eisenthal, Robert; Harrison, Roger

doi:10.1042/bj1450501

Cited by 11 publications

(3 citation statements)

References 15 publications

(18 reference statements)

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“…The high electronegativity of carbon-bound fluorine introduces a polarity more like a C-OH substituent, however, and C-F has been used as a C-OH replacement in carbohydrate derivatives. Briley has used fluorine as an oxygen analogue and has obtained information about the geometric requirements of binding sites and reaction mechanisms involving carbohydrates (Briley et al, 1975).…”

mentioning

confidence: 99%

Stereochemistry of phosphoenolpyruvate carboxylation catalyzed by phosphoenolpyruvate carboxykinase

Hwang¹,

Nowak²

1986

Biochemistry

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The stereochemistry of the carboxylation of phosphoenolpyruvate to yield oxalacetate, catalyzed by chicken liver phosphoenolpyruvate carboxykinase and by Ascaris muscle phosphoenolpyruvate carboxykinase, was determined. The substrate (Z)-3-fluorophosphoenolpyruvate was used for the stereochemical analysis. The carboxylation reaction was coupled to malate dehydrogenase to yield 3-fluoromalate, and the stereochemistry of the products was identified by 19F NMR. In separate experiments, the enantiomeric tautomers of 3-fluorooxalacetate were shown to be utilized by malate dehydrogenase to yield (2R,3R)- and (2R,3S)-3-fluoromalate in nearly identical amounts. The products were identified by 19F NMR. When (Z)-3-fluorophosphoenolpyruvate was used as a substrate for phosphoenolpyruvate carboxykinase from avian liver and from Ascaris, and malate dehydrogenase was used to trap the product, only a single diastereomer was observed. This product was shown to be (2R,3R)-3-fluoromalate in each case. The assignments were based on coupling constants taken from Keck et al. [Keck, R., Hess, H., & Rétey, J. (1980) FEBS Lett. 114, 287]. These results indicate that the stereochemistry of carboxylation, catalyzed by chicken phosphoenolpyruvate carboxykinase and by Ascaris phosphoenolpyruvate carboxykinase, is identical and takes place from the si side of the enzyme-bound phosphoenolpyruvate. The carboxylation reaction was run both in H2O and in D2O. No deuterium incorporation into fluoromalate was shown to occur. The product 3-fluorooxalacetate is thus released from phosphoenolpyruvate carboxykinase as the keto form and is reduced more rapidly by reduced nicotinamide adenine dinucleotide with malate dehydrogenase than by the occurrence of tautomerization.

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confidence: 99%

Stereochemistry of phosphoenolpyruvate carboxylation catalyzed by phosphoenolpyruvate carboxykinase

Hwang¹,

Nowak²

1986

Biochemistry

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“…In family II pyrophosphatases, which belong to the DHH phosphoesterase superfamily (as defined by (5)), the IC 50 for fluoride inhibition was even lower than for NeuTTM (ϳ12 M) (36). In any case fluoride is believed to act as an analog of hydroxide (37). The detailed mechanism of PPP i hydrolysis by NeuTTM requires further investigation, but the lack of effect of vanadate and the strong inhibitory effect of fluoride argue against the existence of a covalent phosphoenzyme intermediate.…”

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confidence: 99%

A Specific Inorganic Triphosphatase from Nitrosomonas europaea

Delvaux

Murty

Gabelica

et al. 2011

Journal of Biological Chemistry

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The CYTH superfamily of proteins is named after its two founding members, the CyaB adenylyl cyclase from Aeromonas hydrophila and the human 25-kDa thiamine triphosphatase. Because these proteins often form a closed ␤-barrel, they are also referred to as triphosphate tunnel metalloenzymes (TTM). Functionally, they are characterized by their ability to bind triphosphorylated substrates and divalent metal ions. These proteins exist in most organisms and catalyze different reactions depending on their origin. Here we investigate structural and catalytic properties of the recombinant TTM protein from Nitrosomonas europaea (NeuTTM), a 19-kDa protein. Crystallographic data show that it crystallizes as a dimer and that, in contrast to other TTM proteins, it has an open ␤-barrel structure. We demonstrate that NeuTTM is a highly specific inorganic triphosphatase, hydrolyzing tripolyphosphate (PPP i ) with high catalytic efficiency in the presence of Mg 2؉ . These data are supported by native mass spectrometry analysis showing that the enzyme binds PPP i (and Mg-PPP i ) with high affinity (K d < 1.5 M), whereas it has a low affinity for ATP or thiamine triphosphate. In contrast to Aeromonas and Yersinia CyaB proteins, NeuTTM has no adenylyl cyclase activity, but it shares several properties with other enzymes of the CYTH superfamily, e.g. heat stability, alkaline pH optimum, and inhibition by Ca 2؉ and Zn 2؉ ions. We suggest a catalytic mechanism involving a catalytic dyad formed by Lys-52 and Tyr-28. The present data provide the first characterization of a new type of phosphohydrolase (unrelated to pyrophosphatases or exopolyphosphatases), able to hydrolyze inorganic triphosphate with high specificity.A large number of phosphohydrolases are able to hydrolyze triphosphorylated compounds, generally nucleoside triphosphates such as ATP or GTP. However, in 2002 we achieved the molecular characterization of a mammalian enzyme that specifically hydrolyzed thiamine triphosphate (ThTP), 6 a compound unrelated to nucleotides (1). This 25-kDa soluble thiamine triphosphatase (ThTPase), which is involved in the regulation of cytosolic ThTP concentrations (2), has near absolute specificity for ThTP (it does not hydrolyze nucleotides) and a high catalytic efficiency. No sequence homology with other known mammalian proteins could be found.Shortly thereafter, Iyer and Aravind (3) observed that the catalytic domains of human 25-kDa ThTPase (1) and CyaB adenylyl cyclase (AC2) from Aeromonas hydrophila (4) define a novel superfamily of domains that, according to these authors, should bind "organic phosphates." This superfamily was called "CYTH" (CyaB-thiamine triphosphatase), and the presence of orthologs was demonstrated in all three superkingdoms of life. This suggested that CYTH is an ancient family of proteins and that a representative must have been present in the last universal common ancestor (LUCA) of all extant life forms. It was proposed (3) that this enzymatic domain might play a central role at the interface between nucleotide a...

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“…Then 30 min later the reaction was stopped with trichloroacetic acid, the mixture was centrifuged and the radioactivity was determined on a 0.1 ml sample of the supernatant. I Fluorinated analogues bear some resemblance to their hydroxylated analogues (Barnett, 1972;Briley et al, 1975). The 3hydroxyglutamic acids were prepared, resolved and incorporated into tripeptides.…”

Section: Carboxylation Of Fluoropeptidesmentioning

confidence: 99%

Vitamin K-dependent carboxylation. Mechanistic studies with 3-fluoroglutamate-containing substrates

Vidal-Cros

Gaudry

Marquet

1990

Biochemical Journal

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The tripeptides t-butyloxycarbonyl-Xaa-Glu-[3H]Val, where Xaa is either (2R,3S)- or (2R,3R)-3-fluoroglutamate (respectively the erythro and the threo isomer), were synthesized and their behaviour during vitamin K-dependent carboxylation was studied. Neither peptide was carboxylated. The erythro compound gave rise to an HF-elimination product representing 1% of the starting material. This HF elimination did not occur during incubation of the threo compound. The formation of the dehydropeptide, probably by elimination of an F- anion from an intermediate carbanion, favours the ionic pathway for vitamin K-dependent carboxylation.

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Fluorine as a hydroxy analogue. Stereospecific phosphorylation of 2-deosy-2-fluoroglycerol by glucerol kinase

Cited by 11 publications

References 15 publications

Stereochemistry of phosphoenolpyruvate carboxylation catalyzed by phosphoenolpyruvate carboxykinase

Stereochemistry of phosphoenolpyruvate carboxylation catalyzed by phosphoenolpyruvate carboxykinase

A Specific Inorganic Triphosphatase from Nitrosomonas europaea

Vitamin K-dependent carboxylation. Mechanistic studies with 3-fluoroglutamate-containing substrates

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