1.
Anal Biochem
; 534: 36-39, 2017 10 01.
Artigo
em Inglês
| MEDLINE
| ID: mdl-28687486
RESUMO
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.