Quantitative extraction and estimation of intracellular nicotinamide nucleotides of Escherichia coli

Lilius, Esa‐Matti; Multanen, Veli-Matti; Toivonen, V.

doi:10.1016/0003-2697(79)90039-3

Cited by 48 publications

(31 citation statements)

References 11 publications

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“…To learn the effects of glucose and ethanol concentration on the metabolism of 2,3,5-trimethyl-1,4-BQ and intracellular NADH concentration, intracellular NADH was extracted and determined according to the method described by Lilius et al (1979). The addition of glucose to yeast cells caused an increase in the oxidative current and intracellular NADH concentration (data not shown).…”

Section: Resultsmentioning

confidence: 98%

Electrochemical Measurement of Yeast Cell Density and Vitality Using 2,3,5-Trimethyl-1,4-benzoquinone and Effect of Ethanol on the Metabolism of Quinone by Yeast Cells

Tsukatani

Ide

Oba

et al. 2003

FSTR

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Section: Resultsmentioning

confidence: 98%

Electrochemical Measurement of Yeast Cell Density and Vitality Using 2,3,5-Trimethyl-1,4-benzoquinone and Effect of Ethanol on the Metabolism of Quinone by Yeast Cells

Tsukatani

Ide

Oba

et al. 2003

FSTR

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“…One is that only methanol:water mixtures were explored. Thus, other solvent compositions [22,26,36,37], pHs [22,28,38,39], or preservatives [37,40,41] may produce substantially superior extracts, with stronger organic solvents potentially having advantages for more completely disrupting membrane structures. The other is that, to gather informative data regarding extraction without introducing complex cell handling procedures or risking metabolite loss via cell leakage, we rely here on the disadvantageously slow approach of harvesting cells by centrifugation.…”

Section: Discussionmentioning

confidence: 99%

Identifying decomposition products in extracts of cellular metabolites

Kimball

Rabinowitz

2006

Analytical Biochemistry

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Most methods of analyzing intracellular metabolites require extraction of metabolites from the cells. A concern in these methods is underestimation of metabolite levels due to incomplete extraction. In comparing extraction methods, it would accordingly seem that the best method for extracting a particular metabolite is the one giving the largest yield. In extracting Escherichia coli with different methanol:water mixtures, we observed that ≥ 50% water gave increased yield of nucleosides and bases compared to ≤ 20% water, as determined by liquid chromatography-tandem mass spectrometry analysis of the resulting extracts. Spiking of the extracts with isotope-labeled nucleotides revealed, however, that the high yield of nucleosides and bases occurred due to decomposition of nucleotides in the water-rich condition, not good extraction. Spiking combined with isotope labeling provides a general approach to detecting decomposition products in extracts of cellular metabolites. For extraction of E. coli with methanol:water, cold temperature and a high methanol fraction minimizes artifacts due to metabolite decomposition.

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“…Determination of NAD+ and NADH was accomplished by the enzymic cycling method of Bernofsky and Swan (1). Extractions of adenine dinucleotides were made according to Lilius et al (8) (Table I). In comparison with the formerly used Potter method, shaking with glass beads generally resulted in a higher number of ruptured grains and a higher yield ofmitochondria plus mitochondrial membranes as deduced from the similarly higher levels of Cyt oxidase activity present in both 15-min and 2-h mitochondria (Table II).…”

Section: Methodsmentioning

confidence: 99%

Isolation-Inflicted Injury to Mitochondria from Fresh Pollen Gradually Overcome by an Active Strengthening during Germination

Hoekstra

Roekel²

1983

Plant Physiol.

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Activities of segments of the electron transport pathway of mitochondria isolated from pollen of Typha latifolia L during the course of germination in vitro were compared with those of mitochondria in intact grains. For this purpose, suitable inhibitors and artificial substrates were selected for their ability to penetrate through the exine, intine, and plasmalemma. In contrast to their counterparts in vivo, mitochondria isolated during the initial stages of germination exhibited low rates of electron transport, resulting from loss of NAD and displacement of cytochrome c from its site of action. The phosphorylative capacity was also impaired. Great caution must be exercised therefore, before interpreting results obtained with isolated mitochondria.The gradually acquired resistance of mitochondria to injury during isolation as germination proceeds was shown to depend on an energyrequiring process and not solely on a rearrangement at the membrane level, or imbibitional differences. Dc novo syntheses of proteins or fatty acids were not required for the strengthening of mitochondria since cycloheximide, chloramphenicol, and cerulenin did not prevent this change. The nature of the energy-requiring process remains obscure. It is probable that strengthening of mitochondrial membranes during seed germination has been misinterpreted due to similar effects of isolational injury.

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Quantitative extraction and estimation of intracellular nicotinamide nucleotides of Escherichia coli

Cited by 48 publications

References 11 publications

Electrochemical Measurement of Yeast Cell Density and Vitality Using 2,3,5-Trimethyl-1,4-benzoquinone and Effect of Ethanol on the Metabolism of Quinone by Yeast Cells

Electrochemical Measurement of Yeast Cell Density and Vitality Using 2,3,5-Trimethyl-1,4-benzoquinone and Effect of Ethanol on the Metabolism of Quinone by Yeast Cells

Identifying decomposition products in extracts of cellular metabolites

Isolation-Inflicted Injury to Mitochondria from Fresh Pollen Gradually Overcome by an Active Strengthening during Germination

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