The Synthesis and Function of Luciferyl-adenylate and Oxyluciferyl-adenylate

Crystallinefirefly luciferase was prepared by the method of Green and McElroy (7).In order to remove traces of contaminating enzymes it was necessary to recrystallize the preparation two or three times.In most of the experiments reported in this paper, three times crystallized enzyme containing about 10 mg. of protein per ml.was used.Crystalline firefly luciferin and oxyluciferin were prepared according to the method of Bitler and McElroy (8).Optical density measurements at 327 rnp and 347 rnp at neutral pH were used to determine the concentration of luciferin and oxyluciferin, respectively.Adenylic acid, ATP, and other nucleotides were obtained from the Sigma Chemical Company.Pabst Laboratories' CoA of approximately 70 per cent purity was used.Reduced CoA was prepared with NaBH., (9).