Active Recombinant Gaussia Luciferase Protein, Streptavidin-Labled

• Cat.No.: Luciferase-105G
• Product Overview: Gaussia(M2)-Luciferase-Streptavidin is a conjugate of one of the brightest luciferases and a protein with the highest known affinity in nature.
  in vitro fusion of two independently expressed proteins in different organisms Gaussia(M2)-Avitag-biotin-Streptavidin

Specification

• Species: Gaussia
• Description: Gaussia Luciferase is a new reporter gene isolated from the marine copepod Gaussia princeps.
• Form: Clear liquid
• Bio-activity: Recommended CTZ concentration for luminometer: 50 μM
  Around 3-fold brighter than wildtyp GLuc-SA
  Binding efficiency greater than 90%
  Very sensitive: use 0.1 to 1 ng of GLuc-SA for biotin coated plates
• Applications: This fusion protein can be used to detect trace amounts of biotinylated protein, DNA, or other biotinylated bio-molecules.
• Storage: Store protein at -80 centigrade for maximum shelf-life, avoid repeated freezing
• Concentration: 1 mg/mL
• Storage Buffer: TBS, pH 7.8 + nonproteinogenic stabilizers
• Dilutions: TBS or Gaussia dilution buffer

Gene Information

• Official Symbol: Luciferase
• Synonyms: Luciferase; Gaussia luciferase; GLuc

Case Study

Case 1: Dijkema FM, et al. Protein Sci. 2024

Gaussia luciferase (GLuc) is one of the most luminescent luciferases known and is widely used as a reporter in biochemistry and cell biology. During catalysis, GLuc undergoes inactivation by irreversible covalent modification. The mechanism by which GLuc generates luminescence and how it becomes inactivated are however not known. Here, researchers show that GLuc unlike other enzymes has an extensively disordered structure with a minimal hydrophobic core and no apparent binding pocket for the main substrate, coelenterazine. From an alanine scan, they identified two Arg residues required for light production. They furthermore show that in addition to coelenterazine, GLuc also can oxidize furimazine, however, in this case without production of light. Both substrates result in the formation of adducts with the enzyme, which eventually leads to enzyme inactivation.

Fig1. Visualization of distance restraints used for the calculation of GLuc's structure.

Fig2. ESI-TOF mass spectrum of wild-type GLuc before (top) and after (bottom) incubation with coelenterazine.

Case 2: Liu Y, et al. Acta Pharm Sin B. 2023

It is discovered that activated caspase-3 tends to induce apoptosis in gasdermin E (GSDME)-deficient cells, but pyroptosis in GSDME-sufficient cells. The high GSDME expression and apoptosis resistance of pancreatic ductal adenocarcinoma (PDAC) cells shed light on another attractive strategy for PDAC treatment by promoting pyroptosis. Here researchers report a hGLuc-hGSDME-PCA system for high-throughput screening of potential GSDME activators against PDAC. This screening system neatly quantifies the oligomerization of GSDME-N to characterize whether pyroptosis occurs under the stimulation of chemotherapy drugs. Based on this system, ponatinib and perifosine are screened out from the FDA-approved anti-cancer drug library containing 106 compounds. Concretely, they exhibit the most potent luminescent activity and cause drastic pyroptosis in PDAC cells.

Fig1. The dose-dependence of GLuc activity in PANC-1 cells.

Fig2. GLuc activity of cells treated with drugs known as caspase-3/GSDME pathway activators or involved in irrelevant pathways.

Application

Gaussia Luciferase (GLuc) is a novel luciferase isolated from Marine organisms with multiple applications.

Bioluminescence detection: GLuc catalyzes the oxidation reaction of the substrate coelenterin and emits light without the involvement of ATP. This property makes it an ideal tool for monitoring biological processes in cells and organisms. GLuc has a high luminous intensity, 1,000 times that of other luciferin enzymes, which makes it very useful for high-sensitivity detection.

Biscistronic expression system: GLuc can be used in the biscistronic expression system to monitor the expression and secretion of biscistronic genes through internal signal peptide-mediated secretion.

Secretion system study: GLuc has secretory signaling peptide, which can be secreted into the extracellular through the endoplasmic reticulum. GLuc can be released from cells through a variety of pathways including classical N-terminal signaling peptide-mediated secretion, internal signaling peptide-mediated secretion, and non-traditional signaling peptide secretion. GLuc's stability and secretion properties make it useful in vivo imaging, for example for monitoring tumor growth and apoptosis.

Protein studies: GLuc can be used as a tool to study oxidative protein folding by monitoring its bioluminescent activity under different oxidative conditions. GLuc can be used for protein interaction analysis to monitor interactions between proteins through the GLuc complementary system.

Fig1. Experimental set up for the secreted Alkaline phosphatase, soluble peptides and Gaussia luciferase assays in body fluids. (Bakhos A Tannous, 2011)