Firefly Luciferase‐based Fusion Proteins and their Applications in Bioanalysis

Smirnova, Daria V.; Ugarova, N. N.

doi:10.1111/php.12656

Cited by 23 publications

(17 citation statements)

References 106 publications

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“…Однако в настоящее время, несмотря на все ограничения, система биолюминесценции светлячков широко используется во многих отраслях науки, и ее практический потенциал еще не исчерпан. Разработанные химерные конструкции, термостабильные, хеморезистентные люциферазы, а также люциферазы с укороченным внутриклеточным периодом полувыведения на основе светлячковой люциферазы описаны в соответствующих обзорах [28,29].…”

Section: люциферазы светлячковunclassified

Palette of Luciferases: Natural Biotools for New Applications in Biomedicine

2020

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Optoanalytical methods based on using genetically encoded bioluminescent enzymes,luciferases, allow one to obtain highly sensitive signals, are non-invasive, and require no external irradiation. Bioluminescence is based on the chemical reaction of oxidation of a low-molecular-weight substrate (luciferin) by atmospheric oxygen, which is catalyzed by an enzyme (luciferase). Relaxation of the luciferin oxidation product from its excited state is accompanied by a release of a quantum of light, which can be detected as an analytical signal.The ability to express luciferase genes in various heterological systems and high quantum yields of luminescence reactions have made these tools rather popular in biology and medicine. Amongseveral naturally available luciferases, a few have been found to be useful for practicalapplication. Luciferase size, the wavelength of its luminescence maximum, enzyme thermostability, optimal pH of the reaction, and the need for cofactors areparameters that may differ for luciferases from different groups of organisms, and this fact directly affects the choice of the application area for each enzyme. It is quite important to overview the whole range of currently available luciferases based ontheir biochemical properties before choosing one bioluminescent probe suitable for a specific application.

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Section: люциферазы светлячковunclassified

Palette of Luciferases: Natural Biotools for New Applications in Biomedicine

2020

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“…The use of bioluminescent proteins as labels provides a high sensitivity of analyte detection in various binding assays over a wide linear range . These properties along with long‐term stability of bioluminescent labels offer their advantages over colorimetric probes.…”

Section: Application Of Copepod Luciferasesmentioning

confidence: 99%

Shining Light on the Secreted Luciferases of Marine Copepods: Current Knowledge and Applications

Markova

Larionova

Vysotski

2019

Photochem & Photobiology

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Copepod luciferases—a family of small secretory proteins of 18.4–24.3 kDa, including a signal peptide—are responsible for bright secreted bioluminescence of some marine copepods. The copepod luciferases use coelenterazine as a substrate to produce blue light in a simple oxidation reaction without any additional cofactors. They do not share sequence or structural similarity with other identified bioluminescent proteins including coelenterazine‐dependent Renilla and Oplophorus luciferases. The small size, strong luminescence activity and high stability, including thermostability, make secreted copepod luciferases very attractive candidates as reporter proteins which are particularly useful for nondisruptive reporter assays and for high‐throughput format. The most known and extensively investigated representatives of this family are the first cloned GpLuc and MLuc luciferases from copepods Gaussia princeps and Metridia longa, respectively. Immediately after cloning, these homologous luciferases were successfully applied as bioluminescent reporters in vivo and in vitro, and since then, the scope of their applications continues to grow. This review is an attempt to systemize and critically evaluate the data scattered through numerous articles regarding the main structural features of copepod luciferases, their luminescent and physicochemical properties. We also review the main trends of their application as bioluminescent reporters in cell and molecular biology.

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“…The availability of luciferase crystal structure of Luciolacruciata and Photinuspyralis has significantly advanced the understanding of key structure-function relationships that account for the efficient enzyme-catalysed emission of light in the firefly [5,6]. Luciferase has the potential to be widely used in molecular biology and bioanalytical systems as a reporter molecule due to the high quantum yield of the bioluminescence, availability of stable mutant forms of the enzyme with prescribed spectral characteristics and abundance of bacterial expression systems suitable for production of recombinant proteins in limitless quantities [7]. Currently, the application of firefly based bioluminescence system is limited to medical and pharmaceutical methods, such as in vivo imaging to visualize tumours, monitoring gene expression and its regulation [8].…”

Section: Introductionmentioning

confidence: 99%

Structural Characterization of Novel Luciferase from Bioluminescence Fungi Verticillium longisporum

Yarima

Sambo

Kwairanga

et al. 2020

BJI

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Luciferase is an enzyme that catalyses a reaction to produce a visible light using an oxidative process, a chemical reaction that is typically referred to as bioluminescent. Insects, bacterial origin or microorganism of marine nature were considered as the mainly sources of discovered luciferase. The protein was commercialized for biomedical and biotechnological use as reporter gene. The first discovered wild form of luciferase originally from Photinu spyralis (firefly). Hence, there is need for both exploration and examination of novel luciferase to be expanded to new sources such as fungal which may likely be exploited to serve commercial purposes and applications. In this study, a novel uncharacterized luciferase protein from a fungal species Verticillium longisporum, was modelled and analysed using bioinformatic tools. The modelled 3D structure is of high quality with a PROCHECK score of 99.5%, ERRAT2 value of 91.01%, and Verify3D score of91.01%, showing that the conformational structure is acceptable. The result showed that the fungal luciferase enzyme share major characteristics with luciferase representative from various fungal and bacterial species. There is only a slight difference in the two nucleotide bindings in V. longisporum with a D/E substitution of D with E and S/T substitution. The difference of the two nucleotides binding from the two proteins may be related to the evolutionary trends. Other differences include increased number of hydrophobic and polar amino acid groups than aromatic and aliphatic ones, as well as more coils and loops with less strands. The distance between the ligand and the binding site that houses Asp 64 and Thr 110 from template proteins (Riboflavin lyaseRcaE) and Asp 543 and Thr 589 from model luciferase is similar. The only difference occurred in the V. longisporum; protein oxidoreductase activities acts on paired donors, incorporate or reduce molecular oxygen, while in the template protein oxidoreductase activities act on single donors with incorporation of molecular oxygen. This study on fungal sourced luciferase present a unique opportunity away from the more well established bacterial and insect based luciferase.

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Firefly Luciferase‐based Fusion Proteins and their Applications in Bioanalysis

Cited by 23 publications

References 106 publications

Palette of Luciferases: Natural Biotools for New Applications in Biomedicine

Palette of Luciferases: Natural Biotools for New Applications in Biomedicine

Shining Light on the Secreted Luciferases of Marine Copepods: Current Knowledge and Applications

Structural Characterization of Novel Luciferase from Bioluminescence Fungi Verticillium longisporum

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