| Literature DB >> 28687486 |
Bruce R Branchini1, Tara L Southworth2, Danielle M Fontaine2, Dawn Kohrt3, Franceine S Welcome2, Catherine M Florentine2, Emma R Henricks2, Demetria B DeBartolo2, Elisa Michelini4, Luca Cevenini4, Aldo Roda4, Martha J Grossel3.
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 ∼2.6-fold higher specific activity. The red-emitting PLR3 was ∼2.5-fold brighter than CBR in living HEK293T and HeLa cells, an improvement consistent with the importance of the Km value in low ATP environments.Entities:
Keywords: ATP; Bioluminescence; Firefly; Imaging; Luciferase; Red-emitting
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Year: 2017 PMID： 28687486 PMCID： PMC5730985 DOI： 10.1016/j.ab.2017.07.001
Source DB: PubMed Journal: Anal Biochem ISSN： 0003-2697 Impact factor: 3.365