By Marlene A. DeLuca (Eds.)
The seriously acclaimed laboratory normal, Methods in Enzymology, is likely one of the so much hugely revered guides within the box of biochemistry. due to the fact 1955, every one quantity has been eagerly awaited, often consulted, and praised by way of researchers and reviewers alike. The sequence includes a lot fabric nonetheless appropriate this day - actually an important booklet for researchers in all fields of lifestyles sciences
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Additional resources for Bioluminescence and Chemiluminescence
Dehydroluciferyl adenylate can be synthesized and purified in an analogous manner. Other Analogs Many additional analogs have been synthesized and have proved to be very useful in probing the molecular mechanism of firefly bioluminescence. A summary of the most useful of these is given in Table II 3,7,12-1~along with references that include descriptions of their synthesis. Of necessity, certain luciferyl analogs have been omitted along with a number of benzothiazole derivatives. However, descriptions of their preparation and properties can generally be found among the references listed, notably references cited in footnotes 5, 7, and 12.
Reaction medium is as described for Fig. 2. KCN (when added) is 2 raM. AdN (when added) is composed of 58/xM AMP and 15 g M ADP. The recording begins with the addition of mitochondria. the intensities of luminescence just before and just after the abrupt change in question. IS] is ATP concentration before the abrupt change and must be determined in a parallel experiment. In the example shown in Fig. 4/~mol of ATP per gram of protein. 1/xM, respectively. These concentrations may be varied to suit differing needs.
C o m m e n t s on the Purification Procedure Table I shows data from a typical purification. Based on the initial light intensity per mol of ATP, the recovery of light-emitting system is about 50%, and the specific activity is increased 6-7 times. The blank values of the two purified preparations listed are equivalent to about [0 -14 tool of ATP. The luciferase is not saturated with iuciferin under the conditions of analysis. Doubling of the luciferin concentration increases the initial light intensity ca.