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High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay
Published on: June 1, 2014
Does the sulfhydryl or the adenine moiety of CoA enhance firefly luciferase activity?
S R Ford1, L M Buck, F R Leach
1Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater 74078-0454, USA.
Biochimica Et Biophysica Acta
|October 25, 1995
Abstract:
Light production by firefly luciferase is limited by product release resulting in flash kinetics. Several compounds (CoA, PPi, and nucleotides) transform the flash-form of light production into continuous light production. The sulfhydryl group of CoA is required; however, since nucleotides are also active, at least two mechanisms (sites) must exist.
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Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.
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Cofactors and Coenzymes
Enzymes are proteins made of amino acids. The functional group of each constituent amino acid catalyzes a wide variety of chemical reactions via ionic interactions or acid-base reactions. However, amino acids cannot catalyze oxidation-reduction and group transfer reactions and need to be aided by non-protein components called cofactors. Cofactors are also referred to as the chemical teeth of an enzyme.
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Cofactors can be metallic ions or organic molecules called coenzymes. These types of helper...

