Determination of the ADP concentration available to participate in energy metabolism in an actin-rich cell, the platelet.

Daniel, James L.; Robkin, Linda; Molish, Ilean R.; Holmsen, Holm

doi:10.1016/s0021-9258(18)36027-7

Cited by 27 publications

(1 citation statement)

References 16 publications

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“…These metabolites were separated by high-voltage paper electrophoresis (60V -cm-1) in 50mM-citrate buffer (Holmsen et al, 1972) and the radioactivity was counted according to standard procedures. As outlined earlier (Daniel et al, 1979(Daniel et al, , 1980, this technique accurately determines metabolically active ATP and ADP and is neither disturbed by actin-bound ADP nor by ATP and ADP stored in the dense granules, which are unavailable for energy metabolism.…”

Section: Assessment Of Energy Consumptionmentioning

confidence: 96%

Quantification of energy consumption in platelets during thrombin-induced aggregation and secretion. Tight coupling between platelet responses and the increment in energy consumption

Verhoeven¹,

Mommersteeg²,

Akkerman³

1984

Biochemical Journal

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The involvement of metabolic energy in platelet responses was investigated by measuring the energy consumption during aggregation and secretion from dense, alpha- and acid-hydrolase-containing granules. Gel-filtered human platelets were stimulated with different amounts of thrombin (0.05-5.0 units X ml-1). At various stages during aggregation and secretion the energy consumption was measured from the changes in metabolic ATP and ADP following abrupt arrest of ATP resynthesis. Stimulation with 5 units of thrombin X ml-1 increased the energy consumption from 6.2 +/- 0.9 to 17.8 +/- 0.4 mumol of ATPeq. X min-1 X (10(11) platelets)-1 during the first 15 s. It decreased thereafter and returned to values found in resting cells after about 30 s. With 0.05 unit of thrombin X ml-1, the energy consumption accelerated more slowly and took at least 3 min before it normalized. A strong positive correlation was found between the velocities of the three secretion responses and the concurrent energy consumption (a) at different stages of the responses induced by a given dose of thrombin, and (b) at different secretion velocities initiated by different amounts of thrombin. When, at different stages of the responses, the extent of secretion was compared with the amount of energy that had been consumed, a strong linear correlation was found with the increment in energy consumption but not with the total energy consumption. This correlation was independent of the concentration of thrombin and indicated that complete secretion from dense, alpha- and acid-hydrolase-containing granules was paralleled by an increment of 4.0, 6.5 and 6.7 mumol of ATPeq. X (10(11) platelets)-1, respectively. An energy cost of 0.7 mumol of ATPeq. X (10(11) platelets)-1 was calculated for separate dense-granule secretion, whereas the combined alpha- and acid-hydrolase granule secretion required 5.3 mumol of ATPeq. X (10(11) platelets)-1. There was no correlation between energy consumption and optical aggregation. In contrast, the rate of single platelet disappearance, which is a measure for the early formation of small aggregates, correlated closely with the rate of energy consumption. Compared with secretion, however, the energy requirement of single platelet disappearance was minor, since 2mM-EDTA completely prevented this response but decreased the energy consumption only slightly. An increase of 0.5-1.0 mumol of ATPeq. X (10(11) platelets)-1 was seen before single platelet disappearance and the three secretion responses were initiated, indicating an increase in energy consuming processes that preceded these responses.(ABSTRACT TRUNCATED AT 400 WORDS)

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Section: Assessment Of Energy Consumptionmentioning

confidence: 96%

Quantification of energy consumption in platelets during thrombin-induced aggregation and secretion. Tight coupling between platelet responses and the increment in energy consumption

Verhoeven¹,

Mommersteeg²,

Akkerman³

1984

Biochemical Journal

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Evaluation of the platelet storage pool deficiency in the feline counterpart of the chediak‐higashi syndrome

et al. 1981

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Cats with the Chediak-Higashi (CH) syndrome have abnormal hemostasis with prolonged bleeding times and normal coagulation times. Platelet aggregation induced by serotonin, ADP, and collagen was impaired. Platelets from normal and CH cats were incubated with 14C-adenine and then gel-filtered. Gel-filtered platelets (GFP) from CH cats contained 63% of the ATP, 38% of the ADP, 100% of the Ca2+, and 75% of the Mg25 of normal platelets. Serotonin could not be detected in CH platelets. Acid hydrolase and total platelet protein of CH platelets was similar to normal platelets. Gel-filtered platelets were treated with thrombin to induce maximal secretion. Secretion of ATP, Ca2+, and Mg2+ was 1.9%, 12.4%, and 16% respectively of normal platelets. ADP secretion by CH platelets was not detectable. The ATP/ADP ratio in the 14C-labeled metabolic pool of normal platelets was similar to that of total measured nucleotide pool of CH platelets. These findings suggest that in feline CH platelets, as in platelets from CH mink and cattle, there is storage pool deficiency that is virtually complete, and the virtual absence of ADP and 5HT may in part account for the abnormal hemostasis. Aggregation of platelets from CH cats was impaired, but these platelets did aggregate to arachidonate, serotonin-induced biphasic aggregation, and the aggregation response to ADP and collagen varied according to the amount of serotonin-induced TxB2 formed. These findings support a major role for arachidonate in platelet activation.

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Compartmentation of Ribonucleotides in PC-12 Cells: Free and Protein Bound Ribonucleotides

Slingerland

Kuilenburg²,

Bodlaender

et al. 1995

Purine and Pyrimidine Metabolism in Man VIII

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Determination of the ADP concentration available to participate in energy metabolism in an actin-rich cell, the platelet.

Cited by 27 publications

References 16 publications

Quantification of energy consumption in platelets during thrombin-induced aggregation and secretion. Tight coupling between platelet responses and the increment in energy consumption

Quantification of energy consumption in platelets during thrombin-induced aggregation and secretion. Tight coupling between platelet responses and the increment in energy consumption

Evaluation of the platelet storage pool deficiency in the feline counterpart of the chediak‐higashi syndrome

Compartmentation of Ribonucleotides in PC-12 Cells: Free and Protein Bound Ribonucleotides

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