Phosphofructokinase: structure and control

Evans, Philip R.; Farrants, George; Hudson, Peter J.

doi:10.1098/rstb.1981.0059

Cited by 167 publications

(59 citation statements)

References 12 publications

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“…The three-dimensional structure ofunliganded E. coli PFK resembles that of the R state (10), indicating that cooperativity (as well as activation by ADP or GDP) could involve only this R state. Also, the PFKs from E. coli and Bacillus stearothermophilus have similar three-dimensional structures, and both undergo a transition from the R to the T state upon binding PEP (5,6,11), but the two enzymes have different saturations by Fru-6P: extremely cooperative for E. coli PFK (3) and hyperbolic for B. stearothermophilus PFK (12). These results indicate that the cooperativity of PFK does not seem coupled to the transition between the crystallographic R and T states.…”

mentioning

confidence: 99%

The cooperativity and allosteric inhibition of Escherichia coli phosphofructokinase depend on the interaction between threonine-125 and ATP.

Auzat

Bras²,

Garel³

1994

Proc. Natl. Acad. Sci. U.S.A.

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During the reaction catalyzed by the phosphofructokinase (EC 2.7.1.11) from Escherichia coli, the phosphoryl group transferred from ATP interacts with [Shir a, Y. & Evans, P. R. (1988) J. Mol. Biol. 204,. The replacement of Thr-125 by serine changes the saturation by fructose 6-phosphate from cooperative to hyperbolic and abolishes the allosteric inhibition by phosphoenolpyruvate. The same changes, a saturation by fructose 6-phosphate that is no longer cooperative and an activity that is no longer inhibited by phosphoenolpyruvate, are observed with wild-type phosphofructokinase when adenosine 5'-[r thio]triphosphate is used instead of ATP as the phosphoryl donor. These two perturbations of the ATP-Thr-125 interaction lead to the suppression of both the allosteric inhibition by phosphoenolpyruvate and the cooperativity of fructose-6-phosphate saturation, as if replacing the neutral oxygen of ATP by sulfur or removing the methyl group of Thr-125 had "locked" phosphofructokinase in its active conformation. The geometry ofthis ATP-Thr-125 interaction and/or the presence of the methyl group on the P-carbon of Thr-125 are crucial for the regulatory properties of phosphofructokinase. This interaction could be a hydrogen bond between the neutral oxygen of the rphosphate of ATP and-the hydroxyl group of Thr-125.The regulation of many biological processes involves allosteric interactions between distinct sites of the same protein, such that ligand binding at one site modifies the functional properties at a distant site. Cooperativity is observed when positive interactions take place between identical sites (1). One ofthe "classic" cooperative enzymes is Escherichia coli phosphofructokinase (PFK; ATP:D-fructose-6-phosphate 1-phosphotransferase, EC 2.7.1.11) (2, 3), which shows a markedly sigmoidal saturation by its substrate fructose 6-phosphate (Fru-6P), with a Hill cooperativity coefficient close to nH = 4 for four Fru-6P sites (3). This enzyme is also sensitive to allosteric effectors, which bind to a regulatory site remote from the active site; PFK is activated by ADP (or GDP) and inhibited by phosphoenolpyruvate (PEP) (3).The steady-state kinetics of E. coli PFK have been interpreted according to the concerted allosteric mechanism (4), in which the protein is in equilibrium between two states, an active R state, which binds Fru-6P and the activator ADP or GDP, and an inactive T state, which binds the inhibitor PEP (3). X-ray crystallography shows that PFK exists in two different conformations, R with activator and substrate (or products) bound (5) and T with inhibitor bound (6), which provides a structural explanation for the allosteric inhibition by PEP (6-8). However, in contrast with hemoglobin for which the differences between the oxy and deoxy states could largely explain the cooperativity of oxygen binding (8,

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mentioning

confidence: 99%

The cooperativity and allosteric inhibition of Escherichia coli phosphofructokinase depend on the interaction between threonine-125 and ATP.

Auzat

Bras²,

Garel³

1994

Proc. Natl. Acad. Sci. U.S.A.

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“…Similarly, our results indicate that each protein examined can multimerize, consistent with previous observations. Indeed, each of these molecules has been shown to form homodimers (DEAD box RNA helicase, enolase, RNase Y, and RnpA [4,6,9,15]), trimers (PNPase [22]), tetramers (phosphofructokinase and RNase J1/J2 [7,23]), or combinations of multimer populations (RNase J1 and J2 form homodimers and homotetramers [17]). While the goal of the current study is to genetically evaluate the potential interacting partners of the S. aureus degradosome, it is clear that such work must be bio- As a preliminary means of validating our bacterial two-hybrid analyses, we biochemically confirmed that purified RNase J1 and RNase J2 interact in vitro; both are hypothesized to be essential S. aureus enzymes (5) and, thus, may play major roles in mRNA decay.…”

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Characterization of Components of the Staphylococcus aureus mRNA Degradosome Holoenzyme-Like Complex

2011

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Bacterial two-hybrid analysis identified the Staphylococcus aureus RNA degradosome-like complex to include RNase J1, RNase J2, RNase Y, polynucleotide phosphorylase (PNPase), enolase, phosphofructokinase, and a DEAD box RNA helicase. Results also revealed that the recently recognized RNase RnpA interacts with the S. aureus degradosome and that this interaction is conserved in other Gram-positive organisms.Escherichia coli bulk RNA degradation is mediated by a holoenzyme complex, the RNA degradosome, which includes the ribonucleases RNase E and polynucleotide phosphorylase (PNPase), an RNA helicase, RhlB, and the glycolytic enzyme enolase (3,20). RNase E is considered the key component of the E. coli degradosome; it catalyzes the initiation of mRNA degradation and serves as a scaffold for the assembly of the other degradosome components (2). By comparison, most Gram-positive bacteria do not contain an RNase E amino acid ortholog and much less is known about their mRNA degradation machinery. Nonetheless, recent studies suggest that the Gram-positive organism Bacillus subtilis produces two RNase E functional orthologs, ribonucleases J1 (RNase J1) and J2 (RNase J2) (17). Using bacterial two-hybrid analyses, both enzymes have been shown to interact with an RNA degradosome-like complex consisting of PNPase, enolase, and a DEAD box RNA helicase (CshA), as well as phosphofructokinase and RNase Y (6).Staphylococcus aureus is a Gram-positive bacterial pathogen of immense health care concern that is evolutionarily similar, but divergent, from the Bacillus genus. Herein we used bacterial two-hybrid and biochemical analyses to measure interactions between putative members of the S. aureus RNA degradosome and compare the results to those with B. subtilis. Results revealed that although many interactions are conserved between both bacterial species, differences are likely to exist. Moreover, we show that the RNase RnpA interacts with components of the S. aureus RNA degradosome and that this interaction is conserved within B. subtilis.Using bacterial two-hybrid analyses, interactions between putative members of the B. subtilis RNA degradosome were recently measured and used to develop the first model of the Gram-positive holoenzyme complex (Fig. 1A) (6, 13). According to the model, the E. coli RNase E orthologs RNase J1 and RNase J2 interact with one another; RNase J1 also interacts with two additional ribonucleases, PNPase and RNase Y, as well as with the glycolytic enzyme phosphofructokinase, each of which binds to the RNA helicase CshA (Fig. 1A). CshA also binds to a second glycolytic enzyme, enolase. Although the mechanism of B. subtilis cellular RNA degradation has yet to be established, RNases J1 and J2 are bifunctional ribonucleases with endonuclease and 5Ј33Ј exoribonuclease activities (8,16,24). In complex, the enzymes act synergistically and may initiate 5Ј mRNA digestion and/or generate endonucleolytic products that could subsequently be digested by RNA helicase-facilitated 3Ј35Ј PNPase exoribonuclease activity (14,1...

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“…These transient interactions of protein complexes can cause several effects such as activation/ inactivation of certain proteins, resulting in the formation of a new binding site. 167,168 Kinetics properties of enzymes can be also altered by denaturation during the entrapment process allowing potential change of the protein specificity to its substrate. [169][170][171] Gaining a clear picture of these basics knowledge will definitely lead to a change of object design to increase the protein load, to control the protein release and to retain the protein integrity and efficacy.…”

Section: Protein Encapsulationmentioning

confidence: 99%

Overview of the main methods used to combine proteins with nanosystems: absorption, bioconjugation, and encapsulation

Marco¹

2009

IJN

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Phosphofructokinase: structure and control

Cited by 167 publications

References 12 publications

The cooperativity and allosteric inhibition of Escherichia coli phosphofructokinase depend on the interaction between threonine-125 and ATP.

The cooperativity and allosteric inhibition of Escherichia coli phosphofructokinase depend on the interaction between threonine-125 and ATP.

Characterization of Components of the Staphylococcus aureus mRNA Degradosome Holoenzyme-Like Complex

Overview of the main methods used to combine proteins with nanosystems: absorption, bioconjugation, and encapsulation

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