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. 1967 Sep;104(3):833–842. doi: 10.1042/bj1040833

Oxidized and reduced nicotinamide–adenine dinucleotide phosphate in tissue suspensions of rat liver

T F Slater 1
PMCID: PMC1271222  PMID: 4383080

Abstract

1. The concentrations of NADP and NADPH2 in homogenates of rat liver (expressed as μg./g. wet wt. of tissue homogenized) were compared with values obtained from intact samples of liver taken from the same female rat. With 0·25m-sucrose alone as the suspending medium, or in combination with tris buffer or 0·01–0·1m-nicotinamide, considerable decreases in the sum of the NADP+NADPH2 concentrations were occasionally observed during 30min. storage of homogenates at 0°. However, addition of 0·5m-nicotinamide+5mm-tris buffer to 0·25m-sucrose for use as a suspending medium maintained the sum of the NADP+NADPH2 concentrations in homogenates at the level found in intact tissue for at least 30min. at 0°. 2. The effects of freezing intact tissue and homogenates in liquid nitrogen before the extraction of NADP and NADPH2 were studied. Freezing alone appears to convert a significant amount (approx. 30%) of liver NADPH2 into an equivalent amount of NADP in intact tissue. This is discussed in terms of the `bound NADP' reported by Burch, Lowry & Von Dippe (1963). 3. The intracellular distributions of NADP and NADPH2 in intracellular fractions of rat liver were studied by using a modified centrifuging scheme that allows extraction of the isolated fractions to be performed within 45min. of killing the animal. Approx. 50% of the total NADP+NADPH2 was found in the large-particle fractions and the remaining 50% was mostly in the soluble fraction of the cell. 4. Further investigations are reported on the nature of `bound NADP' in rat liver. Most of this material appears associated with the `nuclear' (containing nuclei, debris, erythrocytes etc.) or large-mitochondrial fractions, or both, obtained by low-speed centrifuging of rat-liver homogenates. 5. Although in some experiments the variations produced in the concentration of NADPH2 present in large-particle fractions were followed by similar changes in that of `bound NADP', in other cases no such direct relationship was obtained. Addition of phenazine methosulphate, for example, consistently lowered the concentration of NADPH2 yet raised the concentration of `bound NADP' in rat-liver mitochondrial fractions.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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