Coelenterazine-v ligated to Ca2+-triggered coelenterazine- binding protein is a stable and efficient substrate of the red-shifted mutant of Renilla muelleri luciferase : научное издание

Описание

Тип публикации: статья из журнала

Год издания: 2010

Идентификатор DOI: 10.1007/s00216-010-4106-9

Ключевые слова: bioluminescence, calcium, coelenterazine, imaging

Аннотация: 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, thПоказать полностьюe 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 Ca2+-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. © 2010 Springer-Verlag.

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Издание

Журнал: Analytical and Bioanalytical Chemistry

Выпуск журнала: Т. 398, 4

Номера страниц: 1809-1817

ISSN журнала: 16182642

Издатель: Springer-Verlag GmbH

Персоны

  • Stepanyuk G.A. (Photobiology Laboratory,Institute of Biophysics,Russian Academy of Sciences)
  • Malikova N.P. (Photobiology Laboratory,Institute of Biophysics,Russian Academy of Sciences)
  • Markova S.V. (Photobiology Laboratory,Institute of Biophysics,Russian Academy of Sciences)
  • Vysotski E.S. (Photobiology Laboratory,Institute of Biophysics,Russian Academy of Sciences)
  • Unch J. (Promega Biosciences,LLC)
  • Lee J. (Department of Biochemistry and Molecular Biology,University of Georgia)

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