Design and introduction of a disulfide bridge in firefly luciferase: increase of thermostability and decrease of pH sensitivity

2010 ◽  
Vol 9 (8) ◽  
pp. 1167 ◽  
Author(s):  
Mehdi Imani ◽  
Saman Hosseinkhani ◽  
Shahin Ahmadian ◽  
Mahboobeh Nazari
Keyword(s):  
Firefly Luciferase ◽  
Disulfide Bridge ◽  
Ph Sensitivity

Novel application of pH-sensitive firefly luciferases as dual reporter genes for simultaneous ratiometric analysis of intracellular pH and gene expression/location

2014 ◽  
Vol 13 (12) ◽  
pp. 1661-1670 ◽  
Author(s):  
Gabriele V. M. Gabriel ◽  
Vadim R. Viviani
Keyword(s):  
Gene Expression ◽  
Intracellular Ph ◽  
Reporter Genes ◽  
Firefly Luciferase ◽  
Ph Sensitive ◽  
Ph Sensitivity ◽  
Luciferase Gene ◽  
Reporter System ◽  
Dual Reporter

Firefly luciferase pH-sensitivity can be harnessed for ratiometric intracellular pH estimation, providing the first dual reporter system using a single luciferase gene.

Dissection of the molecular mechanisms of protein targeting to the peroxisome using immunofluorescence and immunocryoelectron microscopies

1990 ◽  
Vol 48 (3) ◽  
pp. 44-45
Author(s):  
G-A. Keller ◽  
S. J. Gould ◽  
S. Subramani ◽  
S. Krisans
Keyword(s):  
Mammalian Cells ◽  
Molecular Mechanisms ◽  
Protein Targeting ◽  
Firefly Luciferase ◽  
Peroxisomal Enzyme ◽  
Transfected Cells ◽  
Peroxisomal Targeting ◽  
Targeting Signal ◽  
Series Of Experiments ◽  
Peroxisomal Targeting Signal

Subcellular compartments within eukaryotic cells must each be supplied with unique sets of proteins that must be directed to, and translocated across one or more membranes of the target organelles. This transport is mediated by cis- acting targeting signals present within the imported proteins. The following is a chronological account of a series of experiments designed and carried out in an effort to understand how proteins are targeted to the peroxisomal compartment.-We demonstrated by immunocryoelectron microscopy that the enzyme luciferase is a peroxisomal enzyme in the firefly lantern. -We expressed the cDNA encoding firefly luciferase in mammalian cells and demonstrated by immunofluorescence that the enzyme was transported into the peroxisomes of the transfected cells. -Using deletions, linker insertions, and gene fusion to identify regions of luciferase involved in its transport to the peroxisomes, we demonstrated that luciferase contains a peroxisomal targeting signal (PTS) within its COOH-terminal twelve amino acid.

Sulfonamides Are an Overlooked Class of Electron Donors in Luminogenic Luciferins and Fluorescent Dyes

2018 ◽  
Author(s):  
Deepak K. Sharma ◽  
Spencer T. Adams ◽  
Kate L. Liebmann ◽  
Adam Choi ◽  
Stephen Miller
Keyword(s):  
Fluorescent Dyes ◽  
Bright Light ◽  
Firefly Luciferase ◽  
Electron Donors ◽  
Light Emitting

Many fluorophores, and all bright light-emitting substrates for firefly luciferase, contain hydroxyl or amine electron donors. Here we show that sulfonamides can serve as replacements for these canonical groups. Unlike “caged” carboxamides, sulfonamide analogues enable bioluminescence, and sulfonamidyl luciferins, coumarins, rhodols, and rhodamines are fluorescent in water. Sulfonamide donors thus have previously unappreciated potential to expand the functional repertoire of luminescent molecules.

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