Point mutations in firefly luciferase C-domain demonstrate its significance in green color of bioluminescence

Modestova, Yulia; Koksharov, M. I.; Ugarova, N. N.

doi:10.1016/j.bbapap.2014.04.021

Cited by 16 publications

(12 citation statements)

References 46 publications

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“…Lastly, the side chain carboxylate of Glu451{657} forms a hydrogen bond to the main chain nitrogen of Val465{672} and a salt bridge to the side chain amine of Lys527{743}. An E457K mutation in Luciola mingrelica luciferase at index 657 (Table 2) caused a strong red shift in emission color, and suggested that rigidity in the carboxy-terminal domain is important for green emission in luciferases [63,64]. …”

Section: Resultsmentioning

confidence: 99%

In silico analysis of class I adenylate-forming enzymes reveals family and group-specific conservations

et al. 2018

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Luciferases, aryl- and fatty-acyl CoA synthetases, and non-ribosomal peptide synthetase proteins belong to the class I adenylate-forming enzyme superfamily. The reaction catalyzed by the adenylate-forming enzymes is categorized by a two-step process of adenylation and thioesterification. Although all of these proteins perform a similar two-step process, each family may perform the process to yield completely different results. For example, luciferase proteins perform adenylation and oxidation to produce the green fluorescent light found in fireflies, while fatty-acyl CoA synthetases perform adenylation and thioesterification with coenzyme A to assist in metabolic processes involving fatty acids. This study aligned a total of 374 sequences belonging to the adenylate-forming superfamily. Analysis of the sequences revealed five fully conserved residues throughout all sequences, as well as 78 more residues conserved in at least 60% of sequences aligned. Conserved positions are involved in magnesium and AMP binding and maintaining enzyme structure. Also, ten conserved sequence motifs that included most of the conserved residues were identified. A phylogenetic tree was used to assign sequences into nine different groups. Finally, group entropy analysis identified novel conservations unique to each enzyme group. Common group-specific positions identified in multiple groups include positions critical to coordinating AMP and the CoA-bound product, a position that governs active site shape, and positions that help to maintain enzyme structure through hydrogen bonds and hydrophobic interactions. These positions could serve as excellent targets for future research.

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Section: Resultsmentioning

confidence: 99%

In silico analysis of class I adenylate-forming enzymes reveals family and group-specific conservations

et al. 2018

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“…The resulting spectra are the average of three scans. The molar ellipticity was determined [θ] = (θ × 100 MRW) / (c × l) where θ is the measured ellipticity in degrees at a given wavelength, c is the protein concentration in milligrams per milliliter, and l is the light path length in centimeters [ 27 ].…”

Section: Methodsmentioning

confidence: 99%

SpyTag/SpyCatcher Cyclization Enhances the Thermostability of Firefly Luciferase

Jiang

et al. 2016

PLoS ONE

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SpyTag can spontaneously form a covalent isopeptide bond with its protein partner SpyCatcher. Firefly luciferase from Photinus pyralis was cyclized in vivo by fusing SpyCatcher at the N terminus and SpyTag at the C terminus. Circular LUC was more thermostable and alkali-tolerant than the wild type, without compromising the specific activity. Structural analysis indicated that the cyclized LUC increased the thermodynamic stability of the structure and remained more properly folded at high temperatures when compared with the wild type. We also prepared an N-terminally and C-terminally shortened form of the SpyCatcher protein and cyclization using this truncated form led to even more thermostability than the original form. Our findings suggest that cyclization with SpyTag and SpyCatcher is a promising and effective strategy to enhance thermostability of enzymes.

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“…The Fluc–luciferin system is characterized by a pH‐sensitive BL, in which the emitted light has a peak wavelength at around 560 nm at basic pH, whereas at acidic pH, the wavelength shifts to a maximum at approximately 620 nm . It should be noted that the identity of the light emitter is still a matter of debate, with some researchers claiming the involvement of an anionic keto species and others favoring the presence of the oxyluciferin enolate …”

Section: Molecular Mechanisms Of Chemiluminescent and Bioluminescentmentioning

confidence: 99%

“…[14][15][16][17] It should be noted that the identityo ft he light emitter is still am atter of debate, with some researchers claiming the involvement of an anionic keto species [19,47,67] and others favoring the presence of the oxyluciferin enolate. [21,68,69]…”

Section: Firefly Luciferin-luciferaseblmentioning

confidence: 99%

Chemiluminescence and Bioluminescence as an Excitation Source in the Photodynamic Therapy of Cancer: A Critical Review

2016

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Photodynamic therapy (PDT) of cancer is known for its limited number of side effects, and requires light, oxygen and photosensitizer. However, PDT is limited by poor penetration of light into deeply localized tissues, and the use of external light sources is required. Thus, researchers have been studying ways to improve the effectiveness of this phototherapy and expand it for the treatment of the deepest cancers, by using chemiluminescent or bioluminescent formulations to excite the photosensitizer by intracellular generation of light. The aim of this Minireview is to give a précis of the most important general chemi-/bioluminescence mechanisms and to analyze several studies that apply them for PDT. These studies have demonstrated the potential of utilizing chemi-/bioluminescence as excitation source in the PDT of cancer, besides combining new approaches to overcome the limitations of this mode of treatment.

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Point mutations in firefly luciferase C-domain demonstrate its significance in green color of bioluminescence

Cited by 16 publications

References 46 publications

In silico analysis of class I adenylate-forming enzymes reveals family and group-specific conservations

In silico analysis of class I adenylate-forming enzymes reveals family and group-specific conservations

SpyTag/SpyCatcher Cyclization Enhances the Thermostability of Firefly Luciferase

Chemiluminescence and Bioluminescence as an Excitation Source in the Photodynamic Therapy of Cancer: A Critical Review

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