Analysis of adenosine 3',5'-monophosphate with luciferase luminescence.
TL;DR: A sensitive, relatively simple, assay with wide range linearity has been developed for adenosine 3′,5′-monophosphate (cyclic AMP), which was used to measure the rise in human urine cyclicAMP levels produced by glucagon infusion.
About: This article is published in Analytical Biochemistry.The article was published on 1970-05-01. It has received 95 citations till now. The article focuses on the topics: Adenosine & Adenosine triphosphate.
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TL;DR: Tabulation of adenylate concentrations previously reported for various organisms and tissues supports the prediction, based on enzyme kinetic observations in vitro, that the energy charge is stabilized near 0.85 in intact metabolizing cells of a wide variety of types.
Abstract: The value of the adenylate energy charge, [(adenosine triphosphate) + ½ (adenosine diphosphate)]/[(adenosine triphosphate) + (adenosine diphosphate) + (adenosine monophosphate)], in Escherichia coli cells during growth is about 0.8. During the stationary phase after cessation of growth, or during starvation in carbon-limited cultures, the energy charge declines slowly to a value of about 0.5, and then falls more rapidly. During the slow decline in energy charge, all the cells are capable of forming colonies, but a rapid fall in viability coincides with the steep drop in energy charge. These results suggest that growth can occur only at energy charge values above about 0.8, that viability is maintained at values between 0.8 and 0.5, and that cells die at values below 0.5. Tabulation of adenylate concentrations previously reported for various organisms and tissues supports the prediction, based on enzyme kinetic observations in vitro, that the energy charge is stabilized near 0.85 in intact metabolizing cells of a wide variety of types.
775 citations
TL;DR: The continuous monitoring of ATP concentration by firefly luciferase was used for kinetic determination of enzymes and metabolites and for endpoint analysis of metabolites and was found to be extremely sensitive and convenlent for routine applications.
305 citations
TL;DR: The analytical accuracy of the firefly assay of ATP has been studied and is discussed with special reference to interference with the time course of the bioluminescence, with results shown to result in erroneous assay in a bacterial extract.
234 citations
TL;DR: A rapid, simple, one-step procedure is described for the separation of cyclic AMP from ATP, ADP, AMP, and P 1 based on selective adsorption of all nucleotides except cyclicAMP on hydrous aluminum oxide.
231 citations
TL;DR: Applications of luminescence in the analysis of substancs of clinical interest are surveyed and the advantages, disadvantages, and prospects for luminescent assays are discussed.
Abstract: The various types of chemiluminescent and bioluminescent reactions are described. Applications of luminescence in the analysis of substancs of clinical interest are surveyed. The advantages, disadvantages, and prospects for luminescent assays are discussed.
207 citations
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...Enzyme activity has also been stabilized by bovine serum albumin, sucrose, ascorbic acid, or thioethanol (55, 56)....
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TL;DR: Data indicated a significant role for the enzyme in the control of the levels of adenosine 3’,5’-phosphate present in biological systems, which was very desirable because of the extremely low levels present in most biological materials.
2,056 citations
TL;DR: The evidence presented supports the hypothesis that the positive inotropic response to the catecholamines is mediated by cyclic AMP.
Abstract: Cyclic AMP (adenosine 3',5'-monophosphate or cyclic adenylate) has now been established as a second messenger mediating many of the effects of a variety of hormones. Several of the metabolic effects mediated by cyclic AMP are discussed, and it is suggested that certain other ("functional" or "mechanical") hormonal effects may be similarly mediated. In particular, the evidence presented supports the hypothesis that the positive inotropic response to the catecholamines is mediated by cyclic AMP. Although knowledge of the biological role of cyclic AMP has not been widely applied clinically, sufficient knowledge has now accumulated to make research in this area desirable.
980 citations
563 citations
TL;DR: An enzyme system which catalyzed the formation of cyclic GMP from GTP was demonstrated in small intestine, and this enzyme required Mn++ for maximal activity and was inhibited by ATP, suggesting a rapid turnover of the nucleotide in this tissue.
253 citations