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A Method for Determination of Pyridine Nucleotides Using a Single Extract by Zhiquan Zhang; Jia Yu; Robert C. Stanton is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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Intracellular redox levels play an important role in physiology and pathophysiology. The principal intracellular reductant is NADPH, which is required for both the proper activity of the entire antioxidant system and important prooxidant enzymes such as nitric oxide synthase and NADPH oxidase. Thus an easy and accurate measurement of NADPH is very desirable. The method described in this paper is based on the fact that NADH and NADPH (not NAD ؉ and NADP ؉ ) affect absorbance at 340 nm. A single cell extract is separated into three aliquots (A 1 , A 2 , and A 3 ). A 1 is untreated and the absorbance at 340 nm is measured. A 2 is treated with an enzyme that converts all of the NADP ؉ to NADPH and then the absorbance at 340 nm is measured. A 3 is treated with an enzyme that converts all of the NADPH to NADP ؉ and then the absorbance at 340 nm is measured. A 1 ؊ A 3 is the NADPH content and A 2 ؊ A 1 is the NADP ؉ content of the extract. Using this method, we have obtained full recovery of all added nucleotides from cell extracts, thus making the method suitable for the quick determination of NADP ؉ and NADPH in living cells. We conclude that this method for the measurement of NADP ؉ an

Who reads A Method for Determination of Pyridine Nucleotides Using a Single Extract?

Author

Zhiquan Zhang; Jia Yu; Robert C. Stanton

Publisher

Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 0003-2697)

Published

2000

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)