Bioluminescent measurement in single cardiomyocytes of sudden cytosolic ATP depletion coincident with rigor

Bowers, Keith; Allshire, Ashley; Cobbold, P H

doi:10.1016/0022-2828(92)93159-h

Cited by 70 publications

(33 citation statements)

References 20 publications

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“…Alternative techniques to measure ATP based on firefly luciferase have been developed and used in bacterial and mammalian cells. Luciferase has been injected into mammalian cells to monitor ATP (5,36), and a luciferase reporter system has also been used to monitor ATP in intact cells of E. coli and R. capsulatus (10,14,22,60). The main challenge for this assay in bacterial systems is that cells typically must be treated with permeabilizing drugs such as polymyxin B to allow diffusion of luciferin into cells.…”

Section: Different Extraction Methods Yield Very Different Measuremenmentioning

confidence: 99%

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Effect of Perturbation of ATP Level on the Activity and Regulation of Nitrogenase in Rhodospirillum rubrum

2009

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Nitrogenase activity in Rhodospirillum rubrum and in some other photosynthetic bacteria is regulated in part by the availability of light. This regulation is through a posttranslational modification system that is itself regulated by P II homologs in the cell. P II is one of the most broadly distributed regulatory proteins in nature and directly or indirectly senses nitrogen and carbon signals in the cell. However, its possible role in responding to light availability remains unclear. Because P II binds ATP, we tested the hypothesis that removal of light would affect P II by changing intracellular ATP levels, and this in turn would affect the regulation of nitrogenase activity. This in vivo test involved a variety of different methods for the measurement of ATP, as well as the deliberate perturbation of intracellular ATP levels by chemical and genetic means. To our surprise, we found fairly normal levels of nitrogenase activity and posttranslational regulation of nitrogenase even under conditions of drastically reduced ATP levels. This indicates that low ATP levels have no more than a modest impact on the P II -mediated regulation of NifA activity and on the posttranslational regulation of nitrogenase activity. The relatively high nitrogenase activity also shows that the ATP-dependent electron flux from dinitrogenase reductase to dinitrogenase is also surprisingly insensitive to a depleted ATP level. These in vivo results disprove the simple model of ATP as the key energy signal to P II under these conditions. We currently suppose that the ratio of ADP/ATP might be the relevant signal, as suggested by a number of recent in vitro analyses.The P II family of regulatory proteins is one of the most broadly distributed in nature, being found in all three domains of life (1,20,39,51). P II was originally found to be involved in the regulation of glutamine synthetase (GS) activity by interaction with adenylyltransferase (ATase or GlnE, encoded by glnE) (63, 65). However, P II has subsequently been shown to regulate many other processes involving nitrogen assimilation and metabolism, by directly interacting with different proteins, termed P II receptors (51). In many organisms, more than one P II homolog has been identified. The "housekeeping" homolog is usually referred to as GlnB, encoded by glnB, with other secondary homologs named GlnK, GlnK 2 , GlnJ, GlnY, GlnZ, or Pz (1,12,47,69,78). In a few species of Archaea, a distinct group of P II proteins was named NifI, which is encoded by the nifH-linked gene (39).Because of its physiological diversity, the photosynthetic bacterium Rhodospirillum rubrum is a good model organism in which to study the role of P II . It has three P II homologsGlnB, GlnK, and GlnJ-that play distinct and overlapping functions by interacting with at least six receptors in the cell (71,78,81). Some of these are common to other bacteria, such as GlnE (ATase) that is involved in the regulation for GS activity, the two-component regulatory protein NtrB, and the ammonium transporter (gas channel) A...

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Section: Different Extraction Methods Yield Very Different Measuremenmentioning

confidence: 99%

“…Even a single method performs differently when applied to different organisms (37,46). An alternative technique to measure ATP involves the injection of firefly luciferase into mammalian cells (5,36). The luciferase reporter system has also been used for monitoring ATP in the intact cells of E. coli and R. capsulatus (10,14,22,60).…”

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confidence: 99%

Effect of Perturbation of ATP Level on the Activity and Regulation of Nitrogenase in Rhodospirillum rubrum

2009

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“…Furthermore, in whole mammalian cells, dynamic change of ATP concentration was measured by expression of the recombinant luciferase in vivo. 11,12 The application of this reporter luciferase method to measure a dynamic change of ATP concentration in bacterial cells has been difficult 8,13 because variation in luciferase activity caused by differential transcription, translation, or mRNA/enzyme stability would affect interpretation of the results. And difficulty of luciferin penetration through the bacterial membrane was another problem.…”

Section: Introductionmentioning

confidence: 99%

An Efficient Method for Quantitative Determination of Cellular ATP Synthetic Activity

Hara¹,

Mori²

2006

SLAS Discovery

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The authors have developed an efficient method to measure cellular activity of ATP synthesis. Although ATP is a major energy source of biological reactions, it has been difficult to measure cellular ATP synthetic activity quantitatively. In this report, bioluminescence from the luciferin-luciferase reaction was used for the quantitative measurement. Under the used condition, bioluminescence from standard ATP solution showed no attenuation within several minutes, and the intensity corresponded proportionally to ATP concentrations of the standards. To measure dynamic cellular ATP synthetic activity, combination of osmotic shock and detergent treatment was used to make Escherichia coli cells permeable. ATP was discharged from permeable cells and reacted with externally added luciferase. Because permeable cells used glucose to synthesize and accumulate ATP without further growth, intensity of bioluminescence was increasing during the cellular consumption of glucose. Cellular ATP biosynthetic activity was calculated form the slope of linearly increasing bioluminescence. This permeable cell assay could be applied to high-throughput measuring for dynamic cellular activity of glycolytic ATP synthesis. (Journal of Biomolecular Screening 2006:310-317)

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“…A concept of myocardial ischemia inducing a lag of an early annular mitral motion can generally be proposed. Myocardial relaxation in early phase of diastole is energetically very demanding [22][23][24] . It leads to a diastolic LV dysfunction with increased enddiastolic LV pressures.…”

Section: Discussionmentioning

confidence: 99%

“…An interpretation of such a relationship is not easy, but in the light of experimental and clinical studies considering hemodynamics and diastolic function of LV, it seams that T E-Em is an indirect echocardiographic marker of hibernating myocardial tissue extent. A viable myocardium is partially able to adaptively react to a chronic coronary hypoperfusion state with inhibition of not only an active contraction of the given hibernating segment but also of an energetically demanding early diastolic relaxation of myocardium in the particular region [22][23][24] . From this point of view T E-Em is in our study a single echocardiographic parameter predicting revascularization result in terms of reverse LV remodeling onset.…”

Section: Discussionmentioning

confidence: 99%

Echocardiographic and Cardiac Single Photon Emission Computed Tomography Predictors of Left Ventricle Reverse Remodeling After Surgical Revascularization in Patients With Ischemic Cardiomyopathy and Left Ventricle Systolic Dysfunction

Hutyra¹,

Škála²,

Kamínek³

et al. 2008

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub

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Background:The extent of scar or viable hypocontractile myocardial tissue determines postinfarction left ventricle remodeling. The aim of this pilot study was to evaluate the revascularization eff ect in a group of patients with ischemic cardiomyopathy and LV systolic dysfunction indicated for surgical revascularization, based on evidence for multivessel disease on coronarography and viable myocardium (CMR, SPECT).Aims: To evaluate the revascularization eff ect in patients with ischemic LV systolic dysfunction and to fi nd preoperative predictors of revascularization eff ect.Methods: 33 patients (64±11 years) with baseline LVEF 34.9±9.3 % were included in the study. After a follow-up of 10.7±1.2 months, ECHO and SPECT were performed again. The whole group of patients was divided according to revascularization eff ect (↑LVEF > 5 % and ↓LVESV > 5 % compared with baseline) into revascularization responders (R, n = 22) and nonresponders (NR, n = 11).Results: At baseline there was no diff erence between the subgroups in LVEF (R = 35.7±11.0 % vs. NR = 34.3±8.2 %), EDV (R = 183.6±43.2 vs. NR = 180.2±80.5 ml), ESV (R = 118.5±40.4 vs. NR = 119.7±55.2 ml).The responders showed in a revascularization eff ect subanalysis diff erences in the values of LVEF (+9.8±8.1 %, p < 0.009), reduction of EDV (-39.9±50.9 ml, p = 0.05) and ESV (-35.4±42.6 ml, p = 0,002) compared with baseline.The only preoperative parameters predicting LV reverse remodeling were the T E-Em (R = -10.6±44.1 vs. NR = 29.7±43.7 ms, p = 0.037) and the size of fi xed perfusion defect (FPD) (R = 11.9±13.5 vs. NR = 22.9±15.3 % of LV, p = 0.044).Conclusions: Patients with ischemic LV systolic dysfunction with a preoperatively determined myocardial viability develop LV reverse remodeling. The only preoperative parameters predicting LV reverse remodeling were echocardiographic T E-Em and FPD on SPECT.

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Bioluminescent measurement in single cardiomyocytes of sudden cytosolic ATP depletion coincident with rigor

Cited by 70 publications

References 20 publications

Effect of Perturbation of ATP Level on the Activity and Regulation of Nitrogenase in Rhodospirillum rubrum

Effect of Perturbation of ATP Level on the Activity and Regulation of Nitrogenase in Rhodospirillum rubrum

An Efficient Method for Quantitative Determination of Cellular ATP Synthetic Activity

Echocardiographic and Cardiac Single Photon Emission Computed Tomography Predictors of Left Ventricle Reverse Remodeling After Surgical Revascularization in Patients With Ischemic Cardiomyopathy and Left Ventricle Systolic Dysfunction

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