Sr. Ford et al., EFFECT OF PERIODATE-OXIDIZED ATP AND OTHER NUCLEOTIDES ON FIREFLY LUCIFERASE, Archives of biochemistry and biophysics, 314(2), 1994, pp. 261-267
Addition of periodate-oxidized ATP (oATP) to firefly luciferase-contai
ning reaction mixtures enhanced light production when the reaction mix
ture contained > similar to 8 mu M ATP. The time course of light produ
ction was changed from a flash pattern to a constant light output duri
ng incubations of < similar to 10 min. During longer incubation, firef
ly luciferase was inactivated in a concentration-dependent fashion by
oATP. Firefly luciferase has two different time courses of Light produ
ction that depend on ATP concentration (DeLuca and McElroy, Biochem. B
iophys. Res. Commun,, 123, 764, 1984). The enhancement of light produc
tion occurred only when higher ATP concentrations (> 8 mu M) were used
. There is little effect of oATP on firefly luciferase activity at low
ATP concentrations (< 2 mu M) which gave steady production of light.
ATP did not antagonize the inactivation of firefly luciferase by oATP.
When the oATP was chemically reduced with sodium borohydride (giving
orATP), there was no inactivation of firefly luciferase on incubation.
When orATP was used in a short incubation the enhancement of light pr
oduction and time course change were the same as those observed with o
ATP. The corresponding AMP and adenosine compounds (o and or) were sli
ghtly inhibitory to firefly luciferase activity. ADP was without effec
t but both oADP and orADP enhanced light production. Of these periodat
e-oxidized ADP, AMP, and adenosine derivatives only oADP inactivated f
irefly luciferase. The activating effect can be explained by a change
in the conformation of the enzyme-product complex so that the product
is released faster. In addition there is an inactivation of the enzyme
by certain periodate-oxidized nucleotides during longer incubations,
(C) 1994 Academic Press, Inc.