Anomeric Specificity of the Alkaline Form of Fructose 1,6-Diphosphatase from Rabbit Liver

Benkovic, Patricia A.; Bullard, W P; Maine, Margaret De; Fishbein, Richard; Schray, Keith J.; Steffens, James J.; Benkovic, Stephen J.

doi:10.1016/s0021-9258(19)43020-2

Cited by 30 publications

(15 citation statements)

References 10 publications

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“…In solution, the substrate Fru-1,6-P2 exists mainly in two slowly equilibrating anomeric forms, consisting of about 15% a-anomer and 81% /3-anomer (Midelfort et al, 1976). Rapid quench kinetic studies using Mn2+ as the activating cation showed that the catalytic reaction proceeds in two phases: first a rapid formation of the product, corresponding to about 18% of the available substrate, followed by a slow reaction which has an apparent rate independent of the enzyme concentration and close to the rate of the nonenzymatic conversion of the /3-to a-anomer in solution (Benkovic et al, 1974;Frey et al, 1977). These experiments and the findings that in the crystal structures two Mn2+ and Zn2+ ions are bound in the presence of AhG-l,6-P2 and only one ion is bound in the presence of AhM-l,6-P2 (the analogue of the /3-anomer of Fru-1,6-P2) are consistent with the conclusion that the a-anomer of Fru-1,6-P2 is the true substrate (Zhang et al, 1993).…”

Section: Resultsmentioning

confidence: 99%

Structural aspects of the allosteric inhibition of fructose-1,6-bisphosphatase by AMP: the binding of both the substrate analog 2,5-anhydro-D-glucitol 1,6-bisphosphate and catalytic metal ions monitored by x-ray crystallography

Villeret

Huang²,

Zhang³

et al. 1995

Biochemistry

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The crystal structures of the T form pig kidney fructose-1,6-bisphosphatase (EC 3.1.3.11) complexed with AMP, the substrate analogue 2,5-anhydro-D-glucitol 1,6-bisphosphate (AhG-1,6-P2), and Mn2+ at concentrations of 5, 15, 100, and 300 microM have been determined and refined at resolutions of 2.1-2.3 A to R factors which range from 0.180 to 0.195, respectively. Two metal ions per active site have been identified, one at a binding site of high affinity (metal site 1'), the second in a low affinity site (metal site 2'). The 1-phosphate group of the substrate analogue coordinates to the metal ion at site 1', but not at site 2'. In these four complexes, the distances between the two metal ions are all within 0.2 A of 4.3 A. In the previously determined R form structure of Fru-1,6-Pase complexed with AhG-1,6-P2 and Mn2+, there are also two metal ions in the active site at metal sites 1 and 2. The metal ion at site 1 is only 0.6 A displaced from the metal ion at site 1' in the T form and is also coordinated to the 1-phosphate group of AhG-1,6-P2. However, the second metal ion is located in two distinct sites which are 1.4 A apart in the T and R form structures. In the R form the Mn2+ at site 2 is coordinated to the 1-phosphate group of the substrate analogue. This metal ion is apparently required to orient the phosphate group for nucleophilic attack at the phosphorus center.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Resultsmentioning

confidence: 99%

Structural aspects of the allosteric inhibition of fructose-1,6-bisphosphatase by AMP: the binding of both the substrate analog 2,5-anhydro-D-glucitol 1,6-bisphosphate and catalytic metal ions monitored by x-ray crystallography

Villeret

Huang²,

Zhang³

et al. 1995

Biochemistry

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“…The structure reported here and/or previously published structures may be deadend complexes of FBPase. The kinetic evidence supporting the R-anomer as the substrate (44,46), however, is equally indirect and relates to catalysis by FBPase in the presence of Mn 2+ . Indeed, in the presence of Mg 2+ , data from rapid quenching kinetics infer utilization of both Rand β-anomers of F16P 2 (3).…”

Section: Discussionmentioning

confidence: 99%

Role of a Dynamic Loop in Cation Activation and Allosteric Regulation of Recombinant Porcine Fructose-1,6-bisphosphatase^,

et al. 1998

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A disordered loop (loop 52-72, residues 52-72) in crystal structures of fructose-1,6-bisphosphatase (FBPase) has been implicated in regulatory and catalytic phenomena by studies in directed mutation. A crystal structure of FBPase in a complex with three zinc cations and the products fructose 6-phosphate (F6P) and phosphate (Pi) reveals loop 52-72 for the first time in a well-defined conformation with strong electron density. Loop 52-57 interacts primarily with the active site of its own subunit. Asp68 of the loop hydrogen bonds with Arg276 and a zinc cation located at the putative potassium activation site. Leu56 and Tyr57 of the loop pack against hydrophobic residues from two separate subunits of FBPase. A mechanism of allosteric regulation of catalysis is presented, in which AMP, by binding to its allosteric pocket, displaces loop 52-72 from the active site. Furthermore, the current structure suggests that both the alpha- and beta-anomers of F6P can be substrates in the reverse reaction catalyzed by FBPase. Mechanisms of catalysis are proposed for the reverse reaction in which Asp121 serves as a catalytic base for the alpha-anomer and Glu280 serves as a catalytic base for the beta-anomer.

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“…Through a comparison of the 31P NMR spectra of D-fructose 1,6-bisphosphate, methyl a-D-fructofuranoside 1,6-bisphosphate (11), and methyl ( concluded that the /3-furanose form (6) of D-fructose 1.6-bisphosphate was predominant over the «-form (5) in solution. The 13C NMR spectrum of D-fructose 1.6-bisphosphate also indicated that one major form was present, but the resonances were not assigned.…”

Section: Characterization Of Cyclic Formsmentioning

confidence: 99%

“…The assignments of these resonances were confirmed Figure 7. 31P nuclear magnetic resonance spectrum of D-fructose 1,6-bisphosphate (5,6), pH 6.15. by the synthesis of D-fructose-6,6-d2 1,6-bisphosphate.15'16 In the 31P NMR spectrum of the deuterated analog (Figure 8), the C-6 phosphorus resonances of both furanose forms occur as a broadened singlet at higher field than the triplets of the C-l phosphorus resonances of the a-and d-furanose forms.…”

Section: Characterization Of Cyclic Formsmentioning

confidence: 99%

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Solution structures of the keto sugars and their biologically important phosphate esters

Gray¹

1976

Acc. Chem. Res.

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Carbohydrates, which have long been of interest to chemists as model compounds for the study of the stereochemical aspects of chemical reactions, have recently attracted the interest of biochemists who have examined the stereochemical aspects of their enzyme-catalyzed reactions. A given monosaccharide can exist in several forms in solution, including the acyclic free carbonyl and hydrated carbonyl (gem-diol) forms and the cyclic furanose and pyranose hemiacetal forms. A question arises, however, as to whether enzymes which utilize these sugars as substrates are able to use each of the tautomeric forms. For example, an enzyme may be able to bind all forms but utilize only one in the catalytic reaction. Where only a single form is utilized, the other forms either may be converted to the reactive form by the enzyme or may function as inhibitors. The latter case is especially significant if the reactive form is present in a very low proportion in the tautomeric equilibrium. It is also possible that a given enzyme might bind and utilize only one of the forms present in the equilibrium. It is conceivable that, if the substrate form were used a t a faster rate than it was formed in the tautomeric equilibrium, the rate of the enzyme-catalyzed reaction would be under "tautomeric control".In order to describe the kinetic mechanism of an enzyme whose substrate exists in multiple tautomeric forms in solution, it is necessary to determine the proportion of each form present in the equilibrium and the mode of interaction of each form with the enzyme. Enzymes which utilize phosphorylated esters of keto sugars have received much attention in this regard,1-7 and it has been found that they do indeed recognize and differentially utilize tautomeric forms of their substrates.It is the purpose of this Account to summarize the methods by which the tautomeric compositions of these substrates have been established. The present review will concern itself with the tautomeric compositions of the phosphorylated keto sugars D-fructose 1,6-bisphosphate, D-fructose 6-phosphate, and 1,3-dihydroxy-2-propanone phosphate, and the keto sugars D-fructose, D-tagatose, L-sorbose, and D-pSiC0Se. Although every chemist might not be interested in the structures of these molecules per se, the methods by which these structures have been established are of more general interest because of their applicability to isolation and structural characterization of cell surface antigens and on the development of mycobacterial components as immunotherapeutic agents for cancer.the structural characterization of a variety of organic compounds. Characterization of Acyclic Forms1,3-Dihydroxy-2-propanone phosphate can exist in the acyclic free keto (1) and hydraed keto (2) forms in solution, but D-fructose 1,6-bisphosphate, in addition to the free keto (3) and hydrated keto (4) forms, can also exist in the cyclic a-furanose (5) and @-furanose (6)

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Anomeric Specificity of the Alkaline Form of Fructose 1,6-Diphosphatase from Rabbit Liver

Cited by 30 publications

References 10 publications

Structural aspects of the allosteric inhibition of fructose-1,6-bisphosphatase by AMP: the binding of both the substrate analog 2,5-anhydro-D-glucitol 1,6-bisphosphate and catalytic metal ions monitored by x-ray crystallography

Structural aspects of the allosteric inhibition of fructose-1,6-bisphosphatase by AMP: the binding of both the substrate analog 2,5-anhydro-D-glucitol 1,6-bisphosphate and catalytic metal ions monitored by x-ray crystallography

Role of a Dynamic Loop in Cation Activation and Allosteric Regulation of Recombinant Porcine Fructose-1,6-bisphosphatase^,

Solution structures of the keto sugars and their biologically important phosphate esters

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Anomeric Specificity of the Alkaline Form of Fructose 1,6-Diphosphatase from Rabbit Liver

Cited by 30 publications

References 10 publications

Structural aspects of the allosteric inhibition of fructose-1,6-bisphosphatase by AMP: the binding of both the substrate analog 2,5-anhydro-D-glucitol 1,6-bisphosphate and catalytic metal ions monitored by x-ray crystallography

Structural aspects of the allosteric inhibition of fructose-1,6-bisphosphatase by AMP: the binding of both the substrate analog 2,5-anhydro-D-glucitol 1,6-bisphosphate and catalytic metal ions monitored by x-ray crystallography

Role of a Dynamic Loop in Cation Activation and Allosteric Regulation of Recombinant Porcine Fructose-1,6-bisphosphatase,

Solution structures of the keto sugars and their biologically important phosphate esters

Contact Info

Product

Resources

About

Role of a Dynamic Loop in Cation Activation and Allosteric Regulation of Recombinant Porcine Fructose-1,6-bisphosphatase^,