Red-emitting chimeric firefly luciferase for in vivo imaging in low ATP cellular environments.
Anal Biochem
; 534: 36-39, 2017 10 01.
Article
em En
| MEDLINE
| ID: mdl-28687486
ABSTRACT
Beetle luciferases have been adapted for live cell imaging where bioluminescence is dependent on the cellular availability of ATP, O2, and added luciferin. Previous Photinus pyralis red-emitting variants with high Km values for ATP have performed disappointingly in live cells despite having much higher relative specific activities than enzymes like Click Beetle Red (CBR). We engineered a luciferase variant PLR3 having a Km value for ATP similar to CBR and â¼2.6-fold higher specific activity. The red-emitting PLR3 was â¼2.5-fold brighter than CBR in living HEK293T and HeLa cells, an improvement consistent with the importance of the Km value in low ATP environments.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Trifosfato de Adenosina
/
Luciferases de Vaga-Lume
/
Medições Luminescentes
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Anal Biochem
Ano de publicação:
2017
Tipo de documento:
Article