Synthesis of dehydroluciferin by firefly luciferase: effect of dehydroluciferin, coenzyme A and nucleoside triphosphates on the luminescent reaction.

The formation of dehydroluciferin (L) from luciferin (LH2) in the reaction catalyzed by firefly luciferase (EC 1.13.12.

Time course of luciferyl adenylate synthesis in the firefly luciferase reaction.

The time course of luciferyl adenylate formation in the reaction catalyzed by firefly luciferase (EC 1.13.12.

[The effect of substrates on stability of firefly luciferase, embedded in phosphatidylcholine liposomes].

The effects of substrates (ATP and luciferin) on stability of firefly luciferase embedded into phosphatidylcholine liposomes have been studied. Luciferin did not exert any appreciable influence on enzyme inactivation. Minor concentrations of adenosine 5'-triphosphate destabilized the enzyme; however, the increase in ATP concentration markedly stabilized the enzyme entrapped into liposomes.

New approaches to the preparation and application of firefly luciferase.

Allowing for the lipid nature of firefly luciferase we have developed a new method for obtaining high-activity and high-stability enzyme preparations for bioluminescent microassay. The method includes the step of differential centrifugation in presence of stabilizing additives which entails a partial purification of the enzyme and its essential stabilization likely due to the fact that luciferase retains its lipid environment which plays an important role in catalysis. The resultant luciferase preparation is stable in solution at 4 degrees C for 2-3 months and allows the detection of down to 10(-11) M ATP.

Kinetics of the inactivation of the protein-lipid complex, firefly luciferase, by sodium deoxycholate and its reactivation by phosphatidylcholine.

Firefly luciferase has been shown to be a protein-lipid complex. Phospholipids and neutral lipids bound to luciferase have been identified. Sodium deoxycholate rapidly inactivated the enzyme, but an excess of phosphatidylcholine recovered luciferase activity.