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Thromb Haemost 1985; 53(01): 065-069
DOI: 10.1055/s-0038-1661238
Original Article
Schattauer GmbH Stuttgart

Quantitative and Continuous Analysis of ATP Release from Blood Platelets with Firefly Luciferase Luminescence

Terumasa Higashi
The Department of Physicochemical Physiology, Medical School, Osaka University, Osaka, Japan
,
Akio Isomoto
The Department of Physicochemical Physiology, Medical School, Osaka University, Osaka, Japan
,
Itiro Tyuma
The Department of Physicochemical Physiology, Medical School, Osaka University, Osaka, Japan
,
Eizo Kakishita
*   The Department of Internal Medicine, Hyogo College of Medicine, Hyogo, Japan
,
Michiko Uomoto
*   The Department of Internal Medicine, Hyogo College of Medicine, Hyogo, Japan
,
Kiyoyasu Nagai
*   The Department of Internal Medicine, Hyogo College of Medicine, Hyogo, Japan
› Author Affiliations
Further Information

Publication History

Received 23 December 1984

Accepted 02 November 1984

Publication Date:
18 July 2018 (online)

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Summary

An analytical method was devised to determine the entire course of the adenosine-triphosphate (ATP) release from blood platelets based on a continuous measurement of firefly luciferase luminescence. The equation of the release reaction was derived after due consideration of substrate (ATP) consumption and product (oxyluciferin) inhibition on the luciferase reaction as follows: C, = vt × (Km/Vmax) × (I + It/Ki) + It where Ct is the total concentration of the released ATP at time t, vt is the velocity of the luciferase reaction at time t and is directly measured, It is the concentration of oxyluciferin at time t, and Vmax, Km and Ki are constants. C, of the gel-filtered platelet suspension (GFP) could be determined by substituting vt and It as functions of t. The effects of albumin and temperature on the reaction were also studied.

Key words

ATP release from blood platelets - Firefly luciferase luminescence - Time-course kinetics - Product inhibition
 

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