Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.1.1 (hexokinase)
 5,274 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The outer mitochondrial membrane contains a pore structure which is composed of a 30,000 Da protein, porin. The pore has an internal diameter of 2 nm and exhibits a molecular-sieving exclusion limit between 3000 and 6000 Da. These pores, therefore, provide the exit/entrance port for metabolites moving between mitochondria and the cytosol. Hexokinase binds to porin on the outer surface of mitochondria. The location of hexokinase has evoked a number of theories in which bound hexokinase is given a central role in regulating glycolysis, and, perhaps, the metabolic communication between oxidative and glycolytic metabolism. This is of particular importance in rapidly growing tumor cells in which the aerobic production of lactate and hexokinase activity are highly induced. In the present paper, we summarize the suggested roles of the outer membrane and bound hexokinase in regulation glycolysis of tumor cells. Experiments attempting to elucidate the role of hexokinase binding in the regulation of tumor cell metabolism are presented.
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PMID:The role of the mitochondrial outer membrane in energy metabolism of tumor cells. 242 23

The anomeric specificity of hexokinase was examined in crude homogenates of rat parotid gland, erythrocytes and pancreatic islets. At 8 degrees C, the alpha/beta ratio in maximal velocity averaged 0.73, 0.66 and 0.75 in the parotid, erythrocytes and pancreatic islets, respectively. Hexokinase displayed a greater affinity for alpha- than beta-D-glucose as judged from three criteria: the Km value, the reaction velocity measured with mixtures of the two anomers and their effect upon the phosphorylation of D-[U-14C] glucose in anomeric equilibrium. The latter procedure yielded an alpha/beta ratio in Km close to 0.51, 0.49 and 0.39 in parotid, erythrocytes and pancreatic islets, respectively. Within the limits of this study, the anomeric specificity of mammalian hexokinase would appear to be a mirror image of that of yeast hexokinase.
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PMID:Anomeric specificity of mammalian hexokinase. 243 May 36

The primary cause of red cell destruction in enzymopathies of anaerobic remains controversial and difficult to investigate especially because the erythrocyte population in enzymopenic patients is largely heterogeneous. We have shown that loading human erythrocytes with monospecific enzyme-inactivating antibodies could be useful in understanding the biochemical modifications occurring in enzymopenic erythrocytes and the mechanisms leading to red cell destruction. Hexokinase-inactivating antibodies were prepared and loaded in human erythrocytes using a procedure of encapsulation based on hypotonic hemolysis, isotonic resealing and reannealing. Red blood cells loaded with anti-hexokinase IgG showed 20 +/- 3% residual hexokinase activity while all other enzymes were normal. Lactate production by these cells was 30% of controls while the amount of glucose metabolized in the hexose monophosphate pathway (HMP) was unchanged under resting conditions. However, in the presence of methylene blue HMP rates were only 12% of controls. Determination of adenine nucleotide levels suggests that the antihexokinase-loaded red blood cells are not able to maintain, in vitro, their ATP level as well as their 2,3-diphosphoglycerate. Osmotic fragility, methemoglobin, and reduced glutathione content were near normal. These and other properties of the antihexokinase-loaded erythrocytes were similar to those found in cases of hexokinase deficiency. When the antibody-loaded erythrocytes were chromatrographed on immobilized Protein A columns 66-70% of cells were retained by the column against 0-10% of controls suggesting that hexokinase inactivation promotes autologous IgG binding. Since the phenomenon is known to be associated with red cell phagocytosis, it could be concluded that in hexokinase deficiency red cells are mainly removed by phagocytosis, and that hemolysis probably occurs in cases of oxidative stress when the production of a large amount of reducing equivalents (NADPH) is needed but not provided by the hexokinase-deficient erythrocytes.
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PMID:Human red blood cell loading with hexokinase-inactivating antibodies. An in vitro model for enzyme deficiencies. 250 71

Hexokinase (EC 2.7.1.1) catalyzes the first step in glucose metabolism, using ATP for the phosphorylation of glucose to glucose 6-phosphate. A portion of the HK1 gene was cloned by mixed oligonucleotide primer amplification of cDNA using primers of high complexity. The amino acid sequence for a partial fragment of bovine cardiac muscle HK was determined and used to create primer mixtures of 256- and 1024-fold complexity. Two products were generated from bovine cardiac muscle cDNA which show 82% nucleotide and 93% amino acid identity with a region of rat brain HK1 and cDNA. This work demonstrates that extension and amplification of cDNA probes may be successful even when amino acid sequence data indicate substantial codon degeneracy.
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PMID:Synthesis and characterization of a bovine hexokinase 1 cDNA probe by mixed oligonucleotide primed amplification of cDNA using high complexity primer mixtures. 271 57

Hexokinase-deficient mutants and wild-type Chinese-hamster ovary cells have been used to investigate the role of hexokinase in uptake and accumulation of 2-D-deoxyglucose (2-dGlc). The evidence for a specific sugar transport system in both types of cells is that there is similar saturable phloretin-sensitive uptake of 2-dGlc and 3-O-methyl-D-glucose (3-OMG) in both types of cell. In wild-type cells, 2-dGlc is accumulated to a tissue:medium ratio of 10- and in the mutant only 3-fold; 3-OMG is not accumulated by either mutant or wild-type cells. The evidence that hexokinase affects the membrane transport process is that the rate of exit of free 2-dGlc from wild-type cells is 5-fold less than from mutant cells, whereas there is no difference in the rate of loss of 3-OMG between mutant and wild-type cells.
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PMID:Transport and accumulation of 2-deoxy-D-glucose in wild-type and hexokinase-deficient cultured Chinese-hamster ovary (CHO) cells. 277 78

Hexokinase histochemistry was used to identify brain regions that undergo metabolic changes during the development of hypertension in the spontaneously hypertensive rat (SHR). Photodensitometric measurements of reaction product were made in the commissural subdivision of the nucleus of the tractus solitarius, medial subfornical organ, supraoptic nucleus, magnocellular division of the parvocellular division of the paraventricular nucleus, anterior hypothalamic area, and posterior hypothalamus of prehypertensive SHR (4 weeks) and SHR with developing hypertension (8 weeks). These values were compared with those obtained from age-matched Sprague-Dawley and Wistar-Kyoto rats. At 4 weeks, significantly lower levels of hexokinase were observed in the commissural subdivision of the nucleus of the tractus solitarius and the magnocellular division of the paraventricular nucleus of SHR; a higher level was seen in the posterior hypothalamus. At 8 weeks, significantly higher levels of hexokinase were observed in the anterior hypothalamic area and the posterior hypothalamus. These results can be compared to those from adult SHR where lower levels of activity were found in the parvo- and magnocellular divisions of the paraventricular nucleus. Together these results suggest that, while the role of the magnocellular division of the paraventricular nucleus remains unclear, the commissural subdivision of the nucleus of the tractus solitarius and posterior hypothalamus may participate in the initial events leading to hypertension whereas the parvocellular division of the paraventricular nucleus does not.
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PMID:Metabolic alterations in discrete regions of the rat brain during development of spontaneous hypertension. 280 57

The effects of intracerebroventricular (ICV) colchicine (70 micrograms per rat) on systolic pressures and levels of hexokinase activity in the hypothalamic paraventricular (PVH) and supraoptic (SON) nuclei were investigated in adult normotensive and spontaneously hypertensive rats (SHR). One day after colchicine injection, systolic pressures had dropped significantly in Sprague-Dawley (SD) rats, Wistar-Kyoto (WKY) rats, and SHR; the largest decrease was seen in SHR. Postinjection pressures in SHR were within the normotensive range. No further decreases were observed two days after injections. Quantitative analysis of hexokinase activity in control animals verified that the parvo- and magnocellular PVH (but not SON) of SHR contained significantly lower levels of hexokinase than in WKY or SD rats. Two days after colchicine injection, hexokinase activities in pPVH and mPVH were similar in all three strains. Activity had decreased significantly in SD and WKY rats. In SHR, no differences between control and postinjection values were found. Hexokinase activity in SON was significantly decreased to the same extent in all strains. As metabolic activity in the pPVH, mPVH, and SON decreased after colchicine injection in normotensive rats whereas no such decreases occurred in the pPVH and mPVH of SHR, the findings suggest that colchicine may have differential effects on the metabolic activity of specific cell groups in brain depending on the physiological state of the animal.
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PMID:Effects of colchicine on hexokinase activity in the paraventricular and supraoptic nuclei of spontaneously hypertensive and normotensive rats. 280 10

The role of glucokinase in the regulation of insulin secretion was examined in normal rat pancreatic islets and in chemically- and radiation-induced rat pancreatic B-cell tumours which show an impaired insulin secretory response to glucose. In normal rats glucokinase activity in cytoplasmic fractions of pancreatic islets was decreased with the duration of fasting and increased by refeeding or insulin administration. This observation is consistent with the induction of glucokinase by insulin. Hexokinase activity was only slightly reduced during fasting. Glucokinase activity decreased in cytoplasmic fractions of streptozotocin-nicotinamide-induced rat pancreatic islet cell tumours. Glucokinase activity contributed about 75% to the total glucose phosphorylation capacity in cytoplasmic fractions of normal pancreatic islets and of small (less than 1 mg) streptozotocin-nicotinamide-tumours. This proportion decreased to about 20% in the large streptozotocin-nicotinamide tumours. Glucokinase activity in cytoplasmic fractions of transplantable radiation-induced NEDH (New England Deaconess Hospital) rat B-cell tumours was seven times lower than in normal pancreatic islets and contributed only 15% to the total glucose phosphorylation capacity. In contrast, hexokinase activity of the NEDH tumour B-cells was 2.5 times higher than normal. Decreased glucokinase activity in the chemically- and radiation-induced tumour B-cells appears to result from a loss of the ability of insulin to induce this enzyme and may explain the lack of insulin secretory responsiveness of these tumour B-cells.
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PMID:Defective regulation of glucokinase in rat pancreatic islet cell tumours. 282 Jan 74

Glucose-6-phosphatase (G6Pase) activity was examined cytochemically in the metaphysis of femurs of 3- and 7-day-old rats. G6Pase and hexokinase activities were also examined biochemically in the femur and tibia of 3-day-old animals. The reaction product for G6Pase activity was seen in the endoplasmic reticulum and nuclear envelope of all cell types composing the metaphysis. The amount of the reaction product was abundant in osteoblasts, moderate in osteocytes, and moderate to scarce in osteoclasts and capillary endothelial cells. Biochemical G6Pase activity in the bones was higher than that in the brain, submandibular gland, or pancreas of the animals. Hexokinase activity in the bones was not different from that in the submandibular gland, pancreas, or kidney. The activity ratio of G6Pase and hexokinase in the bones (0.603) was greater than that in the submandibular gland, pancreas, or brain and smaller than that in the kidney. Possible physiological significances of the higher G6Pase activity in osteoblasts are discussed.
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PMID:High glucose-6-phosphatase activity in osteoblasts in the metaphysis of femur of growing rats. 283 86

Yeast hexokinase is a homodimer consisting of two identical subunits. Yeast hexokinase was inactivated by 2-aminothiophenol at 25 degrees C (pH 9.1). The reaction followed pseudo-first-order kinetics until about 70% of the phosphotransferase activity was lost. About 0.65 mol of 2-aminothiophenol/mol of hexokinase was found to be bound after the 70% loss of the enzyme activity. Completely inactivated hexokinase showed a stoichiometry of about 1 mol of 2-aminothiophenol bound/mol of the enzyme. The evidence obtained from kinetic experiments, stoichiometry of the inactivation reaction and fluorescence emission measurements suggested site-site interaction (weak negative co-operativity) during the inactivation reaction. The approximate rate constants for the reversible binding of 2-aminothiophenol to the first subunit (KI) and for the rate of covalent bond formation with only one site occupied (k3) were 150 microM and 0.046 min-1 respectively. The inactivation reaction was pH-dependent. Dithiothreitol, 2-mercaptoethanol and cysteine restored the phosphotransferase activity of the hexokinase after inactivation by 2-aminothiophenol. Sugar substrates protected the enzyme from inactivation more than did the nucleotides. Thus it is concluded that the inactivation of the hexokinase by 2-aminothiophenol was a consequence of a covalent disulphide bond formation between the aminothiol and thiol function at or near the active site of the enzyme. Hexokinase that had been completely inactivated by 2-aminothiophenol reacted with o-phthalaldehyde. Fluorescence emission intensity of the incubation mixture containing 2-aminothiophenol-modified hexokinase and o-phthalaldehyde was one-half of that obtained from an incubation mixture containing hexokinase and o-phthalaldehyde under similar experimental conditions. The intensity and position of the fluorescence emission maximum of the 2-aminothiophenol-modified hexokinase were different from those of the native enzyme, indicating conformational change following modification. Whereas aliphatic aminothiols were completely ineffective, aromatic aminothiols were good inhibitors of the hexokinase. Cyclohexyl mercaptan weakly inhibited the enzyme. Inhibition of the hexokinase by heteroaromatic thiols was dependent on the nature of the heterocyclic ring and position of the thiol-thione equilibrium. The inhibitory function of a thiol is associated with the following structural characteristics: (a) the presence of an aromatic ring, (b) the presence of a free thiol function and (c) the presence of a free amino function in the close proximity of the thiol function.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Inactivation of yeast hexokinase by 2-aminothiophenol. Evidence for a 'half-of-the-sites' mechanism. 284 99


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