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The interaction between neuroglobin (Ngb) and caffeine was studied by spectroscopy. UV-Vis spectra revealed that caffeine did not influence the native hexa-coordinated heme structure of neuroglobin. Fluorescence spectra indicated that caffeine could quench the intrinsic fluorescence of neuroglobin through static quenching mechanisms. The calculated thermodynamic parameters showed that the electrostatic force was the main intermolecular force in the reaction. Results also indicated that the microenvironmental changes of tryptophan and tyrosine residues within neuroglobin are inapparent. Furthermore, the circular dichroism (CD) spectra implied that caffeine could induce the formation of α-helix structure of neuroglobin. 相似文献
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Michele Costanzo Marco Fiocchetti Paolo Ascenzi Maria Marino Marianna Caterino Margherita Ruoppolo 《Molecules (Basel, Switzerland)》2021,26(8)
Neuroglobin (NGB) is a myoglobin-like monomeric globin that is involved in several processes, displaying a pivotal redox-dependent protective role in neuronal and extra-neuronal cells. NGB remarkably exerts its function upon upregulation by NGB inducers, such as 17β-estradiol (E2) and H2O2. However, the molecular bases of NGB’s functions remain undefined, mainly in non-neuronal cancer cells. Human MCF-7 breast cancer cells with a knocked-out (KO) NGB gene obtained using CRISPR/Cas9 technology were analyzed using shotgun label-free quantitative proteomics in comparison with control cells. The differential proteomics experiments were also performed after treatment with E2, H2O2, and E2 + H2O2. All the runs acquired using liquid chromatography–tandem mass spectrometry were elaborated within the same MaxQuant analysis, leading to the quantification of 1872 proteins in the global proteomic dataset. Then, a differentially regulated protein dataset was obtained for each specific treatment. After the proteomic study, multiple bioinformatics analyses were performed to highlight unbalanced pathways and processes. Here, we report the proteomic and bioinformatic investigations concerning the effects on cellular processes of NGB deficiency and cell treatments. Globally, the main processes that were affected were related to the response to stress, cytoskeleton dynamics, apoptosis, and mitochondria-driven pathways. 相似文献
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The fluorescence quenching spectra of neuroglobin by cesium ion, iodide and acrylamide, synchronous fluorescence spectra, and 8-anilino-1-naphthalene-sulfonic acid binding property were investigated. The intrinsic fluorescence of neuroglobin cannot be effectively quenched by cesium ion, with a Stern–Volmer constant of 0.23?±?0.01. However, it can be significantly quenched by iodide and acrylamide with Stern–Volmer constants of 1.30?±?0.04 and 4.56?±?0.10, respectively. The results of synchronous fluorescence spectra indicate that tryptophan residues are distributed on both the surface and interior of neuroglobin molecule, but most of them are buried in the hydrophobic interior. The neuroglobin molecule has different conformational states in various pH solutions. 相似文献
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