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The effect of fructose 1,6 diphosphate on pyruvate kinase from the liver of the flounder (Platichthys flesus L.).

1. Pyruvate kinase purified from flounder liver in two forms, i.e. PKI and PKII, is activated by fructose 1,6 diphosphate. 2. Two or more binding sites for FDP are demonstrated for PKII, the binding to which is influenced by the levels of substrates. 3. FDP reduces or abolishes the cooperative effect of PEP. 4. FDP increases the maximal activity. 5. The inhibition observed at higher levels of ADP is not abolished by FDP.

Adenosine Diphosphate↗

Effects of fructose 2,6-bisphosphate and glucose 1,6-bisphosphate on phosphofructokinase from chicken erythrocytes.

1. Phosphofructokinase (EC 2.7.1.11) from chicken erythrocytes is activated by fructose 2,6-bisphosphate, glucose 1,6-bisphosphate and AMP, and it is inhibited by 2,3-bisphosphoglycerate and inositol hexaphosphate. 2. The stimulatory effects produced by the two bisphosphorylated hexoses are additive and the effects produced by fructose 2,6-bisphosphate and by AMP are synergistic. 3. The activatory effect produced by fructose 2,6-bisphosphate is counteracted by fructose 1,6-bisphosphate. 4. The inhibition produced by both 2,3-bisphosphoglycerate and inositol hexaphosphate is released by fructose 2,6-bisphosphate. 5. It is concluded that, like phosphofructokinase from mammalian tissues, the enzyme from chicken erythrocytes can be modulated by the relative concentrations of those metabolites.

Adenosine Monophosphate↗

Fructose 2,6-bisphosphate in liver of Sparus aurata: influence of nutritional state.

1. Fructose 2,6-bisphosphate (fru-2,6-P2) has been measured in liver and muscle of gilthead sea bream fish, Sparus aurata. 2. The fru-2,6-P2 levels in liver depend on the diet given to the fish: in fish fed a high carbohydrate diet, the fru-2,6-P2 levels are higher than any one previously reported. These changes are associated with differences in the phosphofructokinase 2 activity. 3. Fru-2,6-P2 levels has also been measured in liver of Sparus aurata after different fasting periods. In starved fish, fru-2,6-P2 did not decrease as sharply as in rat. The values found in fish starved for 20 days were similar to those reported for rats that had been starved for 24 hr.

Animals↗

Effects of phosphatase inhibitors on nuclease activity.

Nucleases such as DNase I, which selectively digest chromatin, are inhibited by several commonly used phosphatase inhibitors including sodium bisulfite. Two inhibitors, sodium arsenate and fructose-1,6-diphosphate, did not significantly inhibit nuclease action. Two other effective phosphatase inhibitors, p-chloromercuriphenyl sulfonate and 5,5'-dithiobis(2-nitrobenzoate), can be used during nuclei isolation and then washed out of nuclei before nuclease digestion. Using this procedure, 1mM p-chloromercuriphenyl sulfonate is as effective as 50mM bisulfite in retaining the phosphatase-sensitive mitotic phosphorylations of histones H1 and H3.

4-Chloromercuribenzenesulfonate↗

Fructose-1,6-diphosphatase and glucose-6-phosphatase in newborn rats with intrauterine growth retardation.

The activities of two gluconeogenic enzymes, glucose-6-phosphatase and fructose-1,6-diphosphatase were examined in the normal and intrauterine growth retarded (IUGR) rat during the first 5 days of life. The fructose-1,6-diphosphatase activity, 1.54 +/- 0.10 mumol/min/g liver (means +/- SEM) in control and 1.47 +/- 0.20 in the IUGR rats, increased in both groups on days 2--4 but remained significantly lower in the IUGR rats through day 4 (4.53 +/- 0.6 mumol/min/g liver in control and 3.09 +/- 0.22 mumul/min/g liver in the IUGR rats, P less than 0.01). The glucose-6-phosphatase activity increased similarly in both groups. The weight of the IUGR rats remained lower through the third postnatal day (6.47 +/- 0.42 compared to 8.64 +/- 0.27 g in control rats). Blood glucose concentrations at birth were 117 +/- 11 mg/dl in control rats and 73 +/- 11 mg/dl in the IUGR rats (P less than 0.01). Although the glucose concentrations increased in both groups on days 2--4, the IUGR rats maintained relatively lower levels (P less than 0.01). The results indicate that IUGR fetal rats do not have augmented gluconeogenesis in spite of hypoglycemia. In addition, effective gluconeogenesis in the neonatal period appears to be delayed.

Animals↗

Stereoselectivity of fructose-1,6-bisphosphate aldolase in Thermus caldophilus.

It was recently established that fructose-1,6-bisphosphate (FBP) aldolase (FBA) and tagatose-1,6-bisphosphate (TBP) aldolase (TBA), two class II aldolases, are highly specific for the diastereoselective synthesis of FBP and TBP from glyceraldehyde-3-phosphate (G3P) and dihydroxyacetone phosphate (DHAP), respectively. In this paper, we report on a FBA from the thermophile Thermus caldophilus GK24 (Tca) that produces both FBP and TBP from C(3) substrates. Moreover, the FBP:TBP ratio could be adjusted by manipulating the concentrations of G3P and DHAP. This is the first native FBA known to show dual diastereoselectivity among the FBAs and TBAs characterized thus far. To explain the behavior of this enzyme, the X-ray crystal structure of the Tca FBA in complex with DHAP was determined at 2.2A resolution. It appears that as a result of alteration of five G3P binding residues, the substrate binding cavity of Tca FBA has a greater volume than those in the Escherichia coli FBA-phosphoglycolohydroxamate (PGH) and TBA-PGH complexes. We suggest that this steric difference underlies the difference in the diastereoselectivities of these class II aldolases.

Binding Sites↗

Metabolic and functional effects of carbohydrate substrate with single-dose and multiple-dose potassium cardioplegia.

The isolated working rat heart model of ischemic arrest was used to determine if the addition of carbohydrate substrate to our cardioplegic solution enhanced metabolic and functional myocardial protection. A single-dose cardioplegia technique, as used in earlier studies that showed glucose to have a harmful effect, and a multidose technique similar to that used clinically were studied and compared. Because recent data suggest that fructose-1,6-diphosphate(FDP) may have a protective effect with ischemia, this substrate was also tested and compared to glucose and fructose. In this model, single-dose cardioplegia resulted in poor protection from ischemic injury in all study groups. There was marked improvement in myocardial protection with multidose cardioplegia, and further substantial protection of myocardial function, high-energy phosphate levels, and glycogen stores when carbohydrate substrate was added to the arrest solution. The solution with a higher concentration of glucose (0.5%) provided the best overall metabolic and functional recovery and was clearly superior to fructose and FDP, both of which had about the same protective effect. Improved protection with carbohydrate substrate was accompanied by evidence of substantial increase in glycolytic flux, supporting the idea that increased anaerobic glycolysis can help protect the ischemic myocardium when intermittent reinfusion of cardioplegic solution is done.

Adenosine Triphosphate↗