Hexokinase inhibition

Figure 6-1. The steps of glycolysis. Feedback inhibition of glucose phosphorylation by hexokinase, inhibition of pyruvate kinase, and the main regulatory, rate-limiting step catalyzed by phosphofructoki-nase (PFK-I) are indicated, pyruvate formation and substrate-level phosphorylation are the main outcomes of these reactions. Regeneration of NAD occurs by reduction of pyruvate to lactate during anaerobic glycolysis.

Brewer, G.J., 1969, Erythrocyte metabolism and function Hexokinase inhibition by 2,3-diphosphoglycerate and interaction with ATP and Mgf+, Biochim. Biophys. Acta 192 157. 

Inhibitors of several enzymes in the glycolytic pathway, upon which survival of T. brucei is dependent, have been described. Lonidamine (29) has been shown to inhibit T. brucei hexokinase (IC50 = 850 pM) and be toxic to the parasite (T.b. LD50 = 50 pM) in culture [31]. A series of mannitol derivatives have been discovered, which inhibit T. brucei phos-phofructokinase (TbPFK) [32]. The most potent compound (30) within this series exhibits an IC50 = 23 pM in a recombinant enzyme assay and inhibits parasite growth in vitro (IC50 = 30 pM). 

The role of L-glycerose as a precursor of l sugars is doubtful, since it inhibits glycolysis in plants and animals, probably by the formation of L-sorbose 1-phosphate, which inhibits hexokinase.74-75... 

Hormones have a profound effect on carbohydrate metabolism. Great interest has been aroused by reports of hormonal control of hexokinase activity by specific proteins in animal tissues.99- 100 Hexokinase action is the rate-limiting step in the uptake of D-glucose by muscle. Hexokinase is inhibited in diabetic muscle, but the inhibition can be partially reversed by insulin. A protein fraction from the anterior pituitary gland will inhibit the hexokinase of extracts of brain and muscle, and the effect of this... 

Brain hexokinase is inhibited by its product glucose-6-phosphate and to a lesser extent by adenosine diphosphate 539... 

Brain hexokinase is inhibited by its product glucose-6-phosphate and to a lesser extent by adenosine diphosphate. The isoenzyme of hexokinase found in brain may be soluble in the cytosol or be attached firmly to mitochondria [2 and references therein]. An equilibrium exists between the soluble and the bound enzyme. The binding changes the kinetic properties of hexokinase and its inhibition by Glc-6-P resulting in a more active enzyme. The extent of binding is inversely related to the ATP ADP ratio, i.e. conditions in which energy utilization... 

C7. Crane, R. K., and Sols, A., The non-competitive inhibition of brain hexokinase by glucose-6-phosphate and related compounds. J. Biol. Chem. 210, 597-606 (1954). 

Enzymes are important targets for mercury [71], and sulphydryl-group-containing enzyme being more sensitive to mercuric compounds than a non sulphydryl-group-containing enzyme [72], Enzymes reported to be inhibited include phosphatases [73, 74], dehydrogenases [75,76] and hexokinases [71]. 

Figure 30. A medium complexity model of yeast glycolysis [342], The model consists of nine metabolites and nine reactions. The main regulatory step is the phosphofructokinase (PFK), combined with the hexokinase (HK) reaction into a single reaction vi. As in the minimal model, we only consider the inhibition by its substrate ATP, although PFK is known to have several effectors. External glucose (Glc ) and ethanol (EtOH) are assumed to be constant. Additional abbreviations Glucose (Glc), fructose 1,6 biphosphate (FBP), pool of triosephosphates (TP), 1,3 biphosphogly cerate (BPG), and the pool of pyruvate and acetaldehyde (Pyr).

Treatment of 9-(/ -D-ribofuranosyluronic acid)adenine with diphenylphosphoro-chloridate and orthophosphate or tripolyphosphate yields (62) and (63), which, although unstable, inhibit rabbit AMP aminohydrolase and pyruvate kinase, respectively, with behaviour characteristic of active-site-specific reagents.98 Adenylate kinases from several sources are inactivated by iV6-[2- and 4-fluorobenzoyl]-adenosine-5 -triphosphates, with kinetics characteristic of active-site labelling, although these compounds were without effect on yeast hexokinase and rabbit pyruvate kinase.99... 

Many examples of product inhibition are to found. Some dehydrogenases are inhibited by NADH (a co-product of the reaction), e.g. PDH and isocitrate dehydrogenase (ICD), which are involved with the glycolysis and the TCA cycle are two such examples. Hexokinase isoenzymes in muscle (but not liver) and citrate synthase are inhibited by their products, glucose-6-phosphate and citrate respectively offering a very immediate fine tuning of reaction rate to match cellular requirements and possibly allowing their substrates to be used in alternative pathways. 

A good example of allosteric inhibition is given by hexokinase (HK) isoenzymes of muscle. The product of the HK reaction, glucose-6-P allosterically inhibits the enzyme, so matching the phosphorylation of glucose to its overall metabolism, helps to regulate... 

Competition The rate of transport of one sugar is reduced by the transport of another sugar the kinetics are identical to that of competitive inhibition of hexokinase by the presence of other sugars. 

Specific activation or inhibition Transport of glucose can be increased or decreased by specihc compounds insulin increases the transport whereas phloridzin, a plant glycoside, inhibits glucose transport by muscle. Insulin increases glucokinase activity in liver, whereas a plant sugar, mannoheptulose, inhibits glucokinase activity. Hexokinase is inhibited by its product, glucose 6-phosphate. 

Hexokinase, the activity of which is inhibited by its product, glucose 6-phosphate, which is relieved by phosphate (i.e. phosphate can activate the enzyme when it is inhibited by glncose 6-phosphate). 

Rate experiments that are typically carried out in the presence of different concentrations of an alternative product (or product analog) while using the normal substrates . This approach can be particularly useful when the normal product cannot be used because it is unstable, insoluble, or ineffective (the latter indicated by a very high Ki value). Moreover, the normal product may be consumed as an essential substrate in a coupled assay system for the primary enzyme. Fromm and Zewe used the alternative product inhibition approach in their study of hexokinase. Wratten and Cleland later applied this procedure to exclude the Theorell-Chance mechanism for liver alcohol dehydrogenase. See Abortive Complexes... 

Figure 1. Plot of v/V ax versus the millimolar concentration of total substrate for a model enzyme displaying Michaelis-Menten kinetics with respect to its substrate MA (i.e., metal ion M complexed to otherwise inactive ligand A). The concentrations of free A and MA were calculated assuming a stability constant of 10,000 M k The Michaelis constant for MA and the inhibition constant for free A acting as a competitive inhibitor were both assumed to be 0.5 mM. The ratio v/Vmax was calculated from the Michaelis-Menten equation, taking into account the action of a competitive inhibitor (when present). The upper curve represents the case where the substrate is both A and MA. The middle curve deals with the case where MA is the substrate and where A is not inhibitory. The bottom curve describes the case where MA is the substrate and where A is inhibitory. In this example, [Mfotai = [Afotai at each concentration of A plotted on the abscissa. Note that the bottom two curves are reminiscent of allosteric enzymes, but this false cooperativity arises from changes in the fraction of total "substrate A" that has metal ion bound. For a real example of how brain hexokinase cooperatively was debunked, consult D. L. Purich H. J. Fromm (1972) Biochem. J. 130, 63.

Creatine phosphokinase activity has been reported to be minimally inhibited by hemolysis. Hemoglobin concentrations of 1.25 g/100 ml inhibit 5% and 2.5 g/100 ml, 12% (N5). However, in methods utilizing adenosine diphosphate in the reaction mixture, hemolysates containing 100 mg of hemoglobin per 100 ml may have apparent activities of 5-100 units/liter. The activity is presumably related to adenylate kinase in the erythrocyte (S33). In methods utilizing adenosine diphosphate in a coupled enzyme reaction with hexokinase and glucose-6-phosphatase, the inhibitory effect can be eliminated by adding sufficient adenosine mono-... 

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