860.
It has been shown that the coelenterazine analog, coelenterazine-v, is an efficient substrate for a reaction catalyzed by
Renilla luciferase. The resulting bioluminescence emission maximum is shifted to a longer wavelength up to 40 nm, which allows the
use of some “yellow”
Renilla luciferase mutants for in vivo imaging. However, the utility of coelenterazine-v in small-animal imaging has been hampered
by its instability in solution and in biological tissues. To overcome this drawback, we ligated coelenterazine-v to Ca
2+-triggered coelenterazine-binding protein from
Renilla muelleri, which apparently functions in the organism for stabilizing and protecting coelenterazine from oxidation. The coelenterazine-v
bound within coelenterazine-binding protein has revealed a greater long-term stability at both 4 and 37 °C. In addition, the
coelenterazine-binding protein ligated by coelenterazine-v yields twice the total light over free coelenterazine-v as a substrate
for the red-shifted
R.
muelleri luciferase. These findings suggest the possibility for effective application of coelenterazine-v in various in vitro assays.
相似文献