CdTe quantum dots with green fluorescence generated by bioluminescence resonance energy transfer from aequorin

Jalilian, Nezam; Sajedi, Reza H.; Shanehsaz, Maryam; Gharaat, Morteza

doi:10.1007/s00604-016-2057-3

Cited by 6 publications

(3 citation statements)

References 32 publications

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“…Protein Expression and Purification. First, the apo-aequorin gene (GenBank entry AY601106 36 ) was amplified by Pf u DNA polymerase. The amplified fragment was digested by SalI and XhoI and then cloned into the pPROEX HTB expression vector.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Thermostability of Ctenophore and Coelenterate Ca²⁺-Regulated Apo-photoproteins: A Comparative Study

et al. 2021

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The stabilities of Ca2+-regulated ctenophore and coelenterate apo-photoproteins, apo-mnemiopsin (apo-Mne) and apo-aequorin (apo-Aeq), respectively, were compared biochemically, biophysically, and structurally. Despite high degrees of structural and functional conservation, drastic variations in stability and structural dynamics were found between the two proteins. Irreversible thermoinactivation experiments were performed upon incubation of apo-photoproteins at representative temperatures. The inactivation rate constants (k inact) at 50 °C were determined to be 0.001 and 0.004 min–1 for apo-Mne and apo-Aeq, respectively. Detailed analysis of the inactivation process suggests that the higher thermostability of apo-Mne is due to the higher activation energy (E a) and subsequently higher values of ΔH* and ΔG* at a given temperature. According to molecular dynamics simulation studies, the higher hydrogen bond, electrostatic, and van der Waals energies in apo-Mne can validate the relationship between the thermal adaptation of apo-Mne and the energy barrier for the inactivation process. Our results show that favorable residues for protein thermostability such as hydrophobic, charged, and adopted α-helical structure residues are more frequent in the apo-Mne structure. Although the effect of acrylamide on fluorescence quenching suggests that the local flexibility in regions around Trp and Tyr residues of apo-Aeq is higher than that of apo-Mne, which results in it having a better ability to penetrate acrylamide molecules, the root-mean-square fluctuation of helix A in apo-Mne is higher than that in apo-Aeq. It seems that the greater flexibility of apo-Mne in these regions may be considered as a determining factor, affecting the thermal stability of apo-Mne through a balance between structural rigidity and flexibility.

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Section: ■ Materials and Methodsmentioning

confidence: 99%

Thermostability of Ctenophore and Coelenterate Ca²⁺-Regulated Apo-photoproteins: A Comparative Study

et al. 2021

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“…Following induction of His-tagged protein expression by addition of 1 mM IPTG for 4 h at 20 °C, cells were harvested by centrifugation and lysed by sonication. Afterward, the recombinant protein was purified by Ni-NTA agarose column, as described previously …”

Section: Materials and Methodsmentioning

confidence: 99%

“…Afterward, the recombinant protein was purified by Ni-NTA agarose column, as described previously. 45 Preparation of Semisynthetic Aequorin. To generate semisynthetic aequorin from recombinant apoaequorin, the purified apoaequorin was diluted with 50 mM Tris buffer (pH 7.6) containing 5 mM EDTA (buffer 1).…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

Bioluminescence Detection of Superoxide Anion Using Aequorin

2019

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Although the superoxide anion (O2 –·) is generated during normal cellular respiration and has fundamental roles in a wide range of cellular processes, such as cell proliferation, migration, apoptosis, and homeostasis, its dysregulation is associated with a variety of diseases. Regarding these prominent roles in biological systems, the development of accurate methods for quantification of superoxide anion has attracted tremendous research attention. Here, we evaluated aequorin, a calcium-dependent photoprotein, as a potential bioluminescent reporter protein of superoxide anion. The mechanism is based on the measurement of aequorin bioluminescence, where the lower the concentration of coelenterazine under the oxidation of superoxide anion, the lower the amount aequorin regeneration, leading to a decrease in bioluminescence. The bioluminescence intensity of aequorin was proportional to the concentration of superoxide anion in the range from 4 to 40 000 pM with a detection limit (S/N = 3) of 1.2 pM, which was 5000-fold lower than those of the chemiluminescence methods. The proposed method exhibited high sensitivity and has been successfully applied to the determination of superoxide anion in the plant cell samples. The results could suggest a photoprotein-based bioluminescence system as a highly sensitive, specific, and simple bioluminescent probe for in vitro detection of superoxide anion.

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A dextran mediated multicolor immunochromatographic rapid test strip for visual and instrumental simultaneous detection of Vibrio cholera O1 (Ogawa) and Clostridium botulinum toxin A

Gharaat

Sajedi

Shanehsaz³

et al. 2017

Microchim Acta

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CdTe quantum dots with green fluorescence generated by bioluminescence resonance energy transfer from aequorin

Cited by 6 publications

References 32 publications

Thermostability of Ctenophore and Coelenterate Ca²⁺-Regulated Apo-photoproteins: A Comparative Study

Thermostability of Ctenophore and Coelenterate Ca²⁺-Regulated Apo-photoproteins: A Comparative Study

Bioluminescence Detection of Superoxide Anion Using Aequorin

A dextran mediated multicolor immunochromatographic rapid test strip for visual and instrumental simultaneous detection of Vibrio cholera O1 (Ogawa) and Clostridium botulinum toxin A

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CdTe quantum dots with green fluorescence generated by bioluminescence resonance energy transfer from aequorin

Cited by 6 publications

References 32 publications

Thermostability of Ctenophore and Coelenterate Ca2+-Regulated Apo-photoproteins: A Comparative Study

Thermostability of Ctenophore and Coelenterate Ca2+-Regulated Apo-photoproteins: A Comparative Study

Bioluminescence Detection of Superoxide Anion Using Aequorin

A dextran mediated multicolor immunochromatographic rapid test strip for visual and instrumental simultaneous detection of Vibrio cholera O1 (Ogawa) and Clostridium botulinum toxin A

Contact Info

Product

Resources

About

Thermostability of Ctenophore and Coelenterate Ca²⁺-Regulated Apo-photoproteins: A Comparative Study

Thermostability of Ctenophore and Coelenterate Ca²⁺-Regulated Apo-photoproteins: A Comparative Study