Function of adenosine triphosphate in the activation of luciferin

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Abstract

  • 1.

    1. Using crystalline firefly luciferase and purified luciferin, it is possible to show that light production is associated with the utilization of both luciferin and ATP. The total light produced is directly proportional to the concentration of these two substrates. During luminescence there is a decrease of the 330 mμ absorption peak of luciferin.

  • 2.

    2. The reaction of ATP with luciferin leads to the formation of pyrophosphate and active luciferin (presumably adenylluciferin). The latter compound can either react with oxygen for light production or be hydrolyzed to luciferin and adenylic acid. The latter reaction occurs under anaerobic conditions. Adenylic acid has been identified as one of the products.

  • 3.

    3. The initial reaction of luciferin with ATP is reversible since the addition of PP32 to the reaction mixture leads to the formation of labeled ATP. Luciferin is required for this exchange.

  • 4.

    4. Oxidized luciferin as well as other derivatives of luciferin will replace luciferin as far as pyrophosphate liberation is concerned. In the presence of inorganic pyrophosphatase there is therefore a rapid and complete hydrolysis of ATP to adenylic acid and inorganic phosphate. The results indicate the labile nature of the adenyl compounds.

  • 5.

    5. Adenosine tetraphosphate, uridine triphosphate, cytidine triphosphate, guanosine triphosphate and inosine triphosphate are inactive for both light production and pyrophosphate liberation.

  • 6.

    6. The results are discussed in relation to the mechanism of light emission and the mode of action of pyrophosphate, pyrophosphatase, and coenzyme A on luminescence.

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Contribution No. 150 from the McCollum-Pratt Institute. Supported in part by grant from the National Science Foundation.

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