Phenylalanine ammonia-lyase activity of protoplasts: In vitro inhibition by mannitol and sorbitol

Phenylalanine ammonia-lyase (PAL) activity was strongly inhibited in vitro by D-mannitol and D-sorbitol at concentrations exceeding 50 mM,     

Aldose reductase induced by hyperosmotic stress mediates cardiomyocyte apoptosis: differential effects of sorbitol and mannitol

Cells adapt to hyperosmotic conditions by several mechanisms, including accumulation of sorbitol via induction of the polyol pathway. Failure to adapt to osmotic stress can result in apoptotic cell death. In the present study, we assessed the role of aldose reductase, the key enzyme of the polyol pathway, in cardiac myocyte apoptosis. Hyperosmotic stress, elicited by exposure of cultured rat cardiac myocytes to the nonpermeant solutes sorbitol and mannitol, caused identical cell shrinkage and adaptive hexose uptake stimulation. In contrast, only sorbitol induced the polyol pathway and triggered stress pathways as well as apoptosis-related signaling events. Sorbitol resulted in activation of the extracellular signal-regulated kinase (ERK), p54 c-Jun N-terminal kinase (JNK), and protein kinase B. Furthermore, sorbitol treatment resulting in induction and activation of aldose reductase, decreased expression of the antiapoptotic protein Bcl-xL, increased DNA fragmentation, and glutathione depletion. Apoptosis was attenuated by aldose reductase inhibition with zopolrestat and also by glutathione replenishment with N-acetylcysteine. In conclusion, our data show that hypertonic shrinkage of cardiac myocytes alone is not sufficient to induce cardiac myocyte apoptosis. Hyperosmolarity-induced cell death is sensitive to the nature of the osmolyte and requires induction of aldose reductase as well as a decrease in intracellular glutathione levels.     

Phytochrome-mediated closure of Albizzia lophantha leaflets as affected by sorbitol, mannitol and fusicoccin

The effect of sorbitol and mannitol and their interaction with the effect of fusiccocin has been investigated in phytochrome-mediated nyctinastic closure of Albizzia lophantha leaflets. Treatments were commenced 2 h into the photoperiod. Pairs of leaflets were excised, floated on test solutions, and transferred to darkness immediately after red or far red irradiation. Leaflet angles were measured in darkness for 3 h. Fusicoccin (1 to 100 μ M ) inhibited nyctinastic closure and enhanced reopening in the presence of P_ft (far-red-absorbing form of phytochrome). Fusicoccin effects on far red treated leaflets were not constant and whichever fusicoccin concentration was applied, the effects were small. These results confirm the role of the plasma membrane proton pump in extensor cells during opening. Sorbitol and mannitol (25–100 m M ) increased nyctinastic closure for about 2 h and then enhanced reopening in the presence of P_fr. When sugar alcohols were applied together with fusicoccin, sorbitol and mannitol enhanced the fusicoccin inhibition of closure and a marked synergistic effect was observed. These results indicate that both fusiccocin and the reduction of external osmotic potential caused by sorbitol and mannitol supply may stimulate the activity of the extensor H⁺ pump promoting the reopening.     

Orthophosphate determination in the presence of high concentrations of ATP

A method to measure orthophosphate which contaminates samples of ATP was developed. Concentrations of orthophosphate as low as 0.4% of the ATP concentration were determined using a zinc-molybdate reagent [D. A. Bencini, J. R. Wild, and G. A. O'Donovan, Anal. Biochem. 132, 254-258 (1983)] and continuous spectrophotometric monitoring of chromophore formation. Since the rate of ATP hydrolysis was pseudo-first order and was slow compared to the rate of chromophore formation, the initial concentration of phosphate could be readily determined by extrapolation to zero time. The method is rapid and reproducible, and requires a single, stable reagent.     

Induction of sorbitol dehydrogenase by sorbitol in Aspergillus niger

Summary Evidence is presented for the simultaneous induction of sorbitol dehydrogenase along with fructokinase and repression of glucokinase by sorbitol in Aspergillus niger. Fructose is the first product of sorbitol catabolism.     

Formation of sorbitol 6-phosphate by bovine and human lens aldose reductase, sorbitol dehydrogenase and sorbitol kinase

Formation of sorbitol 6-phosphate by bovine and human lens aldose reductase and sorbitol dehydrogenase by the reduction of glucose 6-phosphate and fructose 6-phosphate, respectively, has been demonstrated. The reaction product has been identified by Dowex-formate column chromatography, gas chromatography and mass spectrometry. Sorbitol 6-phosphate can also be formed by the phosphorylation of sorbitol by lens sorbitol kinase in the presence of ATP.     

Orthophosphate Relations of Root: NO-3 Effects on Orthophosphate Influx, Accumulation and Secretion into the Xylem

Lamaze, T., Sentenac, H. and Grignon, C. 1987. Orthophosphaterelations of root: NO^–₃effects on orthophosphate influx,accumulation and secretion into the xylem.—J. exp. Bot.38: 923–934. Orthophosphate (Pi) accumulation by barley (Hordeum vulgareL.) roots was specifically inhibited by NO^–₃ as comparedto Cl^– and SO₄^{2 –}, and Pi secretion into the xylemwas stimulated. The inhibition of Pi accumulation by NO^–₃was also observed in roots of intact photosynthesizing horsebean(Vicia faba L.), rice (Oryza sativa L.) and soybean (Glycinemax L.) plants. NO^–₃ effects on Pi transport by rootswere more thoroughly investigated with corn (Zea mays L.). Theywere due to intracellular NO^–₃. Pi secretion was stillstimulated by NO^–₃ after Pi withdrawal from the absorptionsolution. ³²Pi influx decreased during Pi accumulation, supportingthe hypothesis that this ion allosterically regulated its owntransport system by feedback control. This control was modulatedby other anions: the decrease was more pronounced in the presenceof nitrate. Chronologically, the depressive effect of NO^–₃on ³²Pi influx appeared after the inhibition of Pi accumulation.Furthermore, under conditions where Pi accumulation was notaffected by NO^–₃, ³²Pi influx and Pi secretion into thexylem became insensitive to the presence of nitrate. Our hypothesisis that the stimulative effect of NO^–₃ on Pi secretionand the depressive one on ³²Pi influx are the repercussionsof an increase in the Pi cytosolic concentration due to an NO^–₃-induced decrease in Pi uptake by the vacuoles. Key words: Root, orthophosphate fluxes, orthophosphate accumulation, nitrate, ionic interaction