Separation of sugar nucleotides, phosphoric esters and free sugars by paper chromatography with solvents containing borates of organic bases

Carminatti, Héctor; Passeron, Susana; Dankert, Marcelo A.; Recondo, Eduardo

doi:10.1016/s0021-9673(01)80372-1

Cited by 70 publications

(12 citation statements)

References 26 publications

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“…Radioactive UDPG was prepared from [U-14C]glucose according to Wright and Robbins [6]. Contaminating uridine diphosphate galactose was separated from UDPG by paper chromatography in morpholininm borate [7]. a-Methyl-glucoside (uniformly labeled with 14C in the glucose moiety) was prepared according to Bollenback 181.…”

Section: Methodsmentioning

confidence: 99%

Action Patterns of Phosphorylase and Glycogen Synthetase on Glycogen

Parodi¹,

Mordoh²,

Krisman³

et al. 1970

European Journal of Biochemistry

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The action patterns of liver and muscle glycogen synthetases and of muscle phosphorylase b on glycogen samples of different molecular weight and on ,&amylase limit dextrins were studied. For this purpose a method for measuring the number of newly added glucose residues that are a t non-reducing ends was developed.It was found that glucose transfer to the non-reducing ends of glycogen catalyzed by liver glycogen synthetase and muscle phosphorylase followed a Poisson distribution.The number of outer chains in the glycogen molecules available to both enzymes appeared to be smaller than the actual number of outer chains in the polysaccharide. The number of such available chains diminished as the glycogen was heavier. For the same glycogen sample, the number of available chains to phosphorylase appeared to be equal or smaller than that to glycogen synthetase.

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Section: Methodsmentioning

confidence: 99%

Action Patterns of Phosphorylase and Glycogen Synthetase on Glycogen

Parodi¹,

Mordoh²,

Krisman³

et al. 1970

European Journal of Biochemistry

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“…The recovery of reducing compounds was 90%. As attempts to resolve these substances in solvents containing borate (7) were not successful, they were measured as a single fraction called "UDP-sugars. "…”

Section: Residual Starchmentioning

confidence: 99%

The Levels of Soluble Nucleotides in Wheat Aleurone Tissue

Collins¹,

Jenner²,

Paleg³

1972

Plant Physiol.

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The content of soluble nucleotides in aleurone layers isolated from mature wheat (Triticum aestivum var. Olympic) grain was investigated. The most abundant nucleotides were adenosine triphosphate, uridine triphosphate, and uridine diphosphoglucose. Smaller amounts of guanosine triphosphate, cytidine triphosphate, adenosine diphosphate, and nicotinamide adenine dinucleotide were also identified. The levels of some of these nucleotides were increased after incubation of the tissue under certain conditions. Nucleotide levels were measured at intervals during incubation of aleurone layers in water. The changes observed are discussed in relation to a response by the tissue to wounding.Gibberellic acid applied to the aleurone layer of mature cereal grain in the absence of the embryo stimulates the activity and secretion of a number of hydrolytic enzymes (10,20,22). A lag phase of some 6 to 8 hr occurs between the addition of GA3 and the first detectable increase in enzyme activity (9, 12).Investigators of the mode of action of GA. have paid close attention to its effect on nucleic acid metabolism (8,25,32 were bisected transversely; and the embryo halves were discarded.The following solvents were redistilled before use: triethylamine, isobutyric acid, ethanol, diethyl ether (over FeSO4 and CaO) and 1-propanol (over KOH). Water was distilled twice and deionized. All other chemicals and solvents used were of reagent grade.All equipment and water was sterilized before use by autoclaving at 121 C and 6.8 kg pressure for 20 min. A sterilizing solution for the half-seeds was prepared by suspending 5 g CaOCl in 100 ml water, shaking the suspension for 10 min, and then filtering. The half-seeds were soaked in the filtrate for 2 hr, rinsed 10 times in water, and then allowed to imbibe water for 24 hr at 30 C.After imbibition, the half-seeds were aseptically transferred to a sterile glass jar; then a weighted polyethylene bottle (sterilized by immersion in 70% ethanol) was inserted in the jar. The jar was capped and rotated at 50 rpm for 30 min (23) with periodic additions of water. The suspension was filtered, and the tissue was returned to the jar and rolled for a further 7 to 8 min. After further filtration and thorough rinsing, the tissue was drained on absorbent paper and weighed out for use. Phillips and Paleg (23) found that bacterial and fungal colonies were almost completely absent even 36 hr after isolation of the tissue. Since most incubations used here were 24 hr or less, no further precautions were taken.The technique used to extract and separate the soluble nucleotides was modified from procedures reported by Cole and Ross (11), Isherwood and Barrett (13), and Jenner (15). Treated tissue was transferred to a cold room, and all operations, unless otherwise stated, were carried out at 4 C. The tissue was first rinsed in ice-cold water and drained on absorbent paper, and then it was disintegrated with an Ultra-Turrax in 5% (w/v) trichloroacetic acid containing 0.15% (w/v) 8-hydroxyquinoline. Each extract was ...

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“…The reaction also was run with labeled UDP-D-xylose as substrate. After 1 hr at 30 C, 3 volumes of 95% ethanol were added to reaction mixtures, and the precipitate was discarded. The supernatant fluid was concentrated and subjected to paper chromatography in solvent 1, and the reaction products were located by ultraviolet quenching or by determination of radioactivity.…”

Section: Resultsmentioning

confidence: 99%