ATP-dependent luminescence

Luciferase-based ATP detection (Promega kinase Glo, Perkin-Elmer Easylite Kinase) ATP-dependent luminescent signal from ludferase conversion of luminal kinase-dependent depletion of ATP is measured Versatile and non-radioactive Signal decrease assay susceptible to luciferase inhibitors Koresawa (2004)... 

STUDY ON ATP-DEPENDENT LUMINESCENCE REACTION OF THE ARM LIGHT ORGANS OF THE LUMINOUS SQUID WA TASEN1A SCINTILLANS... 

In efforts to understand the mechanism involved in the ATP-dependent luminescence, we investigated the conditions for the stabilization, extraction and partial purification of the active insoluble luciferase existing in the arm light organs, and then studied the characteristics of the ATP-dependent luminescence reaction of coelenterazine disulfate in the presence of the active insoluble luciferase obtained. 

Characteristics of ATP-dependent luminescence catalyzed by the arm-organ extract. The concentrations of coelenterazine disulfate, ATP, and MgCI2 have strong effects on the light intensity of luminescence reaction catalyzed by the arm-organ extract. About 8 pM coelenterazine disulfate, 0.5 mM ATP, and 0.8 mM Mg2+ are needed to have maximum levels of luminescence intensity (Fig. 2). 

Function of ATP in the ATP-dependent luminescence reaction catalyzed by the arm-organ extract. The amounts of coelenteramide disulfate, AMP and ADP produced in luminescence reaction under the conditions of standard assay were measured by HPLC. In this experiment, the luminescence reaction mixture had initially contained 4 nmol of coelenterazine disulfate and 250 nmol of ATP, and after 10 min of luminescence reaction, 0.58 nmol of coelenteramide disulfate, 0.1 nmol or less of AMP, and 15 nmol of ADP were found. Thus the molar amount of AMP produced is much less than that of coelenteramide disulfate, indicating that the production of AMP is unrelated to the luminescence reaction. Therefore, the mechanism of ATP-dependent luminescence reaction suggested by Tsuji that involved the formation of adenyl coelenterazine disulfate as a key intermediate, must be incorrect. 

Fig. 4. The [ATP]-dependent steady-state luminescence of oxyluciferin measured as a function of liposome irradiation...

Davenport et al. (1952) were unsuccessful in their attempts to restore the luminescence of the filtered aqueous extract of Luminodesmus. Hastings and Davenport (1957) saw a weak luminescence in their filtered aqueous extracts made from the acetone powder of the millipedes. They found that the luminescence is dependent on pH, with an optimum at about pH 8.9, and that the light intensity could be increased by 10-30% by adding ATP. Hastings and Davenport also measured the luminescence spectrum of live animals, finding an emission peak at 495 nm. 

Numerous methods are available to identify and quantify the extent of mitochondrial dysfunction in hPT cells, such as measurements of ATP concentrations by colorimetric or luminescence-based assays, rates of substrate-dependent... 

All our experiments were carried out on the background of very low ADP concentration (below 10 v>M), which was initially intended to avoid the effect of background luminescence due to the admixture of ATP in ADP preparation. The dependence of the ATP yield on the ADP concentration (Fig. 5) shows that the amount of freshly formed ATP decreases two times when the initial ADP concentration is reduced... 

The europium(III) methacycline complex 19 can be used for spectrofluori-metric determination of lysozyme [49]. The oxytetracycline and doxycycline complexes are both sensitive to ATP in that their luminescence emission is increased in presence of ATP [50, 51]. In addition, complex 17 can be used to determine heparin and NADP in aqueous solution [52,53]. The detection of these biomolecules occurs by means of an enhanced fluorescence emission. Bile acid quenches the luminescence of Eu " doxycycline in a concentration dependent manner [54]. 

Fig. 1 shows a typical trace for the kinetics of luciferase luminescence obtained during illumination with arsenate and ADP. The method used to analyze these curves is also shown. The slow, irreversible phase represents ATP-synthesis supported by endogenous phosphate. The rapid phase in the light, and its reversal in the dark, represent light-dependent synthesis, and non-enzymic hydrolysis, respectively, of ADP-arsenate (Slooten, Nuyten 1983). In the steady state these processes are equally rapid, so that... 

---