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

A comparative study of Mg2+ and Ca2+ effects on the ability of rat skeletal muscle hexokinase isozyme II to bind mitochondrial membranes isolated from the same source was carried out. It was found that the binding ability of the enzyme increases in a similar way in the presence of equimolar amounts of both cations. The dependence of binding ability on cation concentration is hyperbolic, which points to the existence of specific and equivalent metal binding sites during hexokinase attachment to the membranes. Substitution of Ca2+ for Mg2+ does not influence the tightness of the enzyme binding to membranes, which can be evidenced from the type of dependence of the bound hexokinase solubilization degree on KCl concentration in the eluting buffer. The enzyme absorption mediated by various cations is accompanied by corresponding changes in its kinetic properties (V, Km for glucose, Ki for ADP). The role of bivalent cations in the formation of the specific hexokinase-membrane binding is discussed.
 ...
PMID:[The role of bivalent cations in the binding of hexokinase II isoenzyme to mitochondrial membranes]. 370 21

It was found that in the presence of Mg2+ (pH 7.5) rat skeletal muscle hexokinase isozyme II is firmly adsorbed on mitochondrial and artificial phospholipid membranes (lecithin liposomes). In both cases the adsorption isotherm has similar quantitative and qualitative characteristics, which points to the absence of specific binding sites on the membranes. Under these conditions, immobilization of hexokinase on various membranes is concomitant with similar changes in the enzyme stability upon storage as well as with the pH-dependence of the enzyme activity. It was demonstrated that the bound hexokinase form has a greater value of V, an increased affinity for glucose and a decreased sensitivity to the inhibitory action of glucose-6-phosphate as compared to the free form. Besides, this form is in a greater degree subjected to the inhibitory influence of ADP with respect to glucose. In this case, the enzyme affinity for ATP and the Ki value for ADP with respect to ATP is practically the same both for the free and membrane-bound forms. The data obtained suggest that the phospholipid component of mitochondrial membranes participates in the enzyme binding in the presence of Mg2+. It was assumed that the model system used in the present study, i.e., hexokinase-Mg2+-liposomes, may be successfully used for the analysis of an adsorption mechanism of regulation of hexokinase activity in the cell.
 ...
PMID:[Interaction of hexokinase II isoenzyme from rat skeletal muscles with lecithin liposomes]. 373 Apr 44

The mechanism of retraction of the longitudinal flagellum of Ceratium tripos was studied by making extracted models of the flagellum. Non-detergent models extracted in low ionic strength medium containing 1 M-glucose, 10 mM-EDTA, and 50 mM-Tris X HCl buffer (pH 8.0), retracted when Ca2+, Mg2+, Ba2+, Sr2+, Mn2+ or Cd2+ was applied locally with a glass capillary. A demembranated model of the flagellum was made with an extraction medium containing 0.8-1.0 M-glucose, 20 mM-Tris-acetate (pH 7.8), 2 mM-EGTA, 5-7 mM-MgSO4, 0.1 M-potassium glutamate and 0.1% Triton X-100. The model required a concentration of Mg2+ of a few mmol/l for successful reactivation of both retraction and undulation, and about 0.1 M-potassium glutamate (or sodium glutamate) for reactivation of undulation. Neither type of motion of the models could be reactivated above 35 degrees C. Ca2+ induced the retraction at pCa 5.5 or less. In addition to Ca2+, Mn2+, Ba2+, Sr2+ and Cd2+ also induced retraction but Mg2+, La3+ or Tb3+ did not. Although ATP was required for undulation, it was not required for retraction. Co-incubation with hexokinase to remove contaminating ATP did not suppress the retraction. The potent ATPase inhibitor, orthovanadate, inhibited undulation at 10 micron but did not inhibit retraction even at 2 mM. SH blockers, N-ethylmaleimide and dithio-bis-nitrobenzoic acid strongly suppressed undulation but had no effect on retraction. Calmodulin inhibitors, trifluoperazine and chlorpromazine, also had no effect on retraction. These data indicate that undulation is generated by a 9 + 2 microtubular axoneme using energy released by hydrolysis of ATP and that retraction can be induced by Ca2+ without a requirement for ATP.
 ...
PMID:Extraction model of the longitudinal flagellum of Ceratium tripos (Dinoflagellida): reactivation of flagellar retraction. 385 92

We describe a simple method for determining magnesium in serum by using hexokinase (EC 2.7.1.1) and glucose-6-phosphate dehydrogenase (EC 1.1.1.49). The method is based on determination of the reaction rate of hexokinase activated by Mg2+, which participates in the hexokinase reaction as the substrate in the form of a Mg X ATP2- complex. The reaction rate is determined from the change in absorbance at 340 nm as NADPH is produced by the action of glucose-6-phosphate dehydrogenase. This simple and rapid spectrophotometric method does not require expensive instrumentation, but results correlate satisfactorily with those obtained by atomic absorption spectroscopy. Thus, the present method gives a "true" value for magnesium in serum, a value appreciably lower than that obtained by an earlier colorimetric method, the Xylidyl Blue II method (Biochem Med 7: 208-217, 1973), which lacks specificity.
 ...
PMID:Direct spectrophotometry of magnesium in serum after reaction with hexokinase and glucose-6-phosphate dehydrogenase. 398 98

1. Magnesium ions are the most effective bivalent ions in the glucokinase reaction. 2. The molecular weight of rat hepatic glucokinase is 48000-49000 as assessed by gel filtration on Sephadex G-100. 3. Anomalous kinetic behaviour at low glucose concentrations appears to be due to the formation during the purification procedure of fragments possessing modified catalytic properties, but is unlikely to be of physiological significance. 4. Extension of previous studies (Parry & Walker, 1966) suggests that glucokinase catalyses a reaction of the random Bi Bi type similar to that of yeast hexokinase. 5. The inhibitory effects of various thiol reagents suggest that a thiol group may be involved at or near the binding site of the acceptor molecule.
 ...
PMID:Further properties and possibel mechanism of action of adenosine 5'-triphosphate-D-glucose 6-phosphotransferase from rat liver. 558 91

The regulation of extramicrosomal Ca2+ concentration maintained by suspensions of rat insulinoma microsomes was studied using Ca2+-selective minielectrodes. The Ca2+-transporting activity was MgATP dependent and correlated with the endoplasmic reticulum marker NADPH-cytochrome c reductase. When incubated in a high KCl medium containing Mg2+ and phosphate, the microsomes lowered [Ca2+] within less than 10 min to around 0.2 microM. They had a high Ca2+-sequestering activity since they were able to take up and retain several small Ca2+ additions. No evidence for a Na+/Ca2+ countertransport was obtained. The accumulated Ca2+ was released by the Ca2+ ionophore A23187 or upon transforming ATP into ADP using glucose plus hexokinase. The addition of ADP, at concentrations present in cells, resulted in a dose-dependent and reversible net Ca2+ efflux from the microsomes until a higher [Ca2+] steady state was reached. This was specific for ADP since GDP, UDP, CDP, IDP, and the nonhydrolyzable analogue methylene-ADP as well as AMP and cAMP did not reproduce the effect. Insulin secretory granules were unable to lower medium [Ca2+] or to take up a pulse addition of Ca2+. However, most of the large granular calcium content was released by A23187. The addition of Na+ and lowering or increasing medium pH by 0.2 pH unit did not induce Ca2+ uptake or efflux from the secretory granules. The results indicate that insulinoma endoplasmic reticulum but not insulin secretory granules may play a critical role in the regulation of cytosolic Ca2+. A variation in cellular ADP content following secretagogue addition might modulate Ca2+ fluxes across the endoplasmic reticulum and contribute in raising cytosolic Ca2+.
 ...
PMID:Regulation of Ca2+ transport by isolated organelles of a rat insulinoma. Studies with endoplasmic reticulum and secretory granules. 608 82

The kinetic properties of the glycosomal hexokinase (HG)2 and phosphofructokinase (PFK) from Trypanosoma brucei bloodstream forms were investigated. Hexokinase has a very high affinity for glucose (Km = 17 microM) and exhibits a broad pH optimum with a maximum at pH 7.8. No indications have been found for regulation of HK activity. Phosphofructokinase behaves as an allosteric protein with respect to its substrate, fructose-6-phosphate. 5'-AMP acts as a positive allosteric effector. The apparent Km for 5'-AMP is extremely low (7 microM). The other substrate for PFK is Mg2+-ATP chelate which activates the enzyme in a hyperbolic manner. Excess of ATP over Mg2+ is inhibitory. The enzyme needs Mg2+ for full activity. Compounds known to be positive or negative heterotrophic modifiers of PFK in other organisms are without effect. It is concluded that PFK and HK probably do not play a regulatory role in glycolysis in T. brucei.
 ...
PMID:Regulation of glycolysis in Trypanosoma brucei: hexokinase and phosphofructokinase activity. 612 64

Deciliation of Paramecium tetraurelia by a Ca2+ shock procedure releases a discrete set of proteins which represent about 1% of the total cell protein. Marker enzymes for cytoplasm (hexokinase), endoplasmic reticulum (glucose-6-phosphatase), peroxisomes (catalase), and lysosomes (acid phosphatase) were not released by this treatment. Among the proteins selectively released is a Ca2+-dependent ATPase. This enzyme has a broad substrate specificity which includes GTP, ATP, and UTP, and it can be activated by Ca2+, Sr2+, or Ba2+, but not by Mg2+ or by monovalent cations. The crude enzyme has a specific activity of 2-3 mumol/min per mg; the optimal pH for activity is 7.5. ATPase, GTPase, and UTPase all reside in the same protein, which is inhibited by ruthenium red, is irreversibly denatured at 50 degrees C, and which has a sedimentation coefficient of 8-10 S. This enzyme is compared with other surface-derived ATPases of ciliated protozoans, and its possible roles are discussed.
 ...
PMID:A Ca2+-activated ATPase specifically released by Ca2+ shock from Paramecium tetraurelia. 612 13

A cation-sensing electrode can be used to monitor enzymatic reactions if their substrates and products differ in the ability to bind the cation. In the first method, the electrode potential vs time curve is recorded. The second, metal-stat method consists of measuring the rate at which substrate must be added to the reaction medium in order to keep the electrode potential at a constant level. These approaches have been used to assay inorganic pyrophosphatase, carboxypeptidase A, and hexokinase with the Mg2+ and Cu2+ ions as the indicators.
 ...
PMID:Use of cation-selective electrodes in enzyme assays. 613 61

Mg2+-ATPase activity was identified in the cytosol of human erythrocytes. A partial purification of this activity was achieved by an initial DEAE-Sephadex column chromatography, followed by gel filtration on Sephadex G-100 and then a second DEAE-Sephadex chromatography procedure. The enzyme appeared in the void volume of the Sephadex G-100 column and was retained on an Amicon XM100A ultrafiltration membrane. The molecular weight of the enzyme was estimated to be 113 000 from SD gels. The above purification protocol yielded an enzyme with an optimal pH between 7.6 and 8.2. The enzyme activity increased linearly between 30 and 44 degrees C. It was stable for several months at -20 degrees C. Magnesium was essential for activity, but the rate attainable with Mn2+ was at least as great as that due to Mg2+. No other divalent cation was able to substitute for Mg2+ or Mn2+. Neither low nor high Ca2+ concentrations significantly affected the enzymatic activity. Substrate specificity studies showed that ATP was the preferred substrate followed by CTP (46% of the rate produced by ATP). Hydrolysis of GTP, UTP, ITP and ADP was less than 10% of the rate seen with ATP. No phosphatase, pyrophosphatase, phosphodiesterase, hexokinase, phosphofructokinase or adenylate cyclase activity could be detected in this enzyme preparation. Calmodulin, which stimulates the (Ca2+ + Mg2+)-ATPase of the human erythrocyte membrane, failed to enhance the Mg2+-ATPase activity. Of considerable interest, the activity of this Mg2+-ATPase was enhanced approximately 5-fold by low concentrations of mercuric ion, p-hydroxymercuribenzoate and DTNB, but was much less sensitive to iodoacetamide.
 ...
PMID:Partial purification and characterization of a novel Mg2+-dependent ATPase present in the cytosol from human erythrocytes. 615 Jul 30


<< Previous 1 2 3 4 5 6 7 8 9 Next >>