Comparative Study of Toluidine Blue O and Methylene Blue Binding to Lysozyme and Their Inhibitory Effects on Protein Aggregation

Saha, Baishakhi; Chowdhury, Sourav; Sanyal, Dwipanjan; Chattopadhyay, Krishnananda; Kumar, Gopinatha Suresh

doi:10.1021/acsomega.7b01991

Cited by 50 publications

(37 citation statements)

References 77 publications

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“…As for BPB, most of these dyes show static quenching of fluorescence with a 1:1 stoichiometry, which implies a single binding site that must be located next to some of the aromatic residues responsible for the intrinsic fluorescence of the protein (Baugher et al, 1974). Recent studies on the binding of the dyes toluidine blue O and methylene blue by isothermal titration calorimetry (ITC) also corroborated a single binding site, which was modelled next to tryptophan residues 62 and 63 (Saha et al, 2018). However, our crystallographic structures show that the binding of BPB is not specific and we have found more than one BPB molecule attached to the surface of the protein.…”

Section: Discussionmentioning

confidence: 93%

Lysozyme crystals dyed with bromophenol blue: where has the dye gone?

Plaza-Garrido

Salinas-Garcia

Alba-Elena

et al. 2020

Acta Cryst Sect D Struct Biol

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Protein crystals can easily be coloured by adding dyes to their mother liquor, but most structures of these protein–dye complexes remain unsolved. Here, structures of lysozyme in complex with bromophenol blue obtained by soaking orthorhombic and tetragonal crystals in a saturated solution of the dye at different pH values from 5.0 to 7.5 are reported. Two different binding sites can be found in the lysozyme–bromophenol blue crystals: binding site I is located near the amino- and carboxyl-termini, while binding site II is located adjacent to helices α1 (residues 4–15) and α3 (residues 88–100). In the orthorhombic crystals soaked at pH 7.0, binding of the dye takes place in both sites without significant changes in the unit cell. However, soaking tetragonal crystals with bromophenol blue results in two different complexes. Crystals soaked at pH 5.5 (HEWL-T1) show a single dye molecule bound to site II, and the crystals belong to space group P43212 without significant changes in the unit cell (a = b = 78.50, c = 37.34 Å). On the other hand, crystals soaked at pH 6.5 in the presence of imidazole (HEWL-T2) show up to eight molecules of the dye bound to site II, and display changes in space group (P212121) and unit cell (a = 38.00, b = 76.65, c = 84.86 Å). In all of the structures, the dye molecules are placed at the surface of the protein near to positively charged residues accessible through the main solvent channels of the crystal. Differences in the arrangement of the dye molecules at the surface of the protein suggest that the binding is not specific and is mainly driven by electrostatic interactions.

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Section: Discussionmentioning

confidence: 93%

Lysozyme crystals dyed with bromophenol blue: where has the dye gone?

Plaza-Garrido

Salinas-Garcia

Alba-Elena

et al. 2020

Acta Cryst Sect D Struct Biol

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“…When applied topically, MB improved fibroblast proliferation as well as collagen and elastin fibers production in tegument [34]. At the mitochondrial level, MB acts on the first, third, and fourth respiratory complex.…”

Section: Methylene Bluementioning

confidence: 92%

“…Concentration from 0.05 µM to 1 M of MB solutions [32,33] MB is the first synthetic dye ever used as an antiseptic in clinical therapy [11]. This effect is granted by its capacity to stain the nucleic acids; moreover, MB is a photosensitizing agent for photodynamic inactivation of RNA viruses including human immunodeficiency virus (HIV), hepatitis B virus, and hepatitis C virus in plasma, while oxidative damage to isolated DNA caused by exposure to UV light is minimal in humans [34].…”

Section: Preoperative and Intraoperative Use In Cardiac Surgerymentioning

confidence: 99%

Comparative Study Regarding the Properties of Methylene Blue and Proflavine and Their Optimal Concentrations for In Vitro and In Vivo Applications

et al. 2020

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Methylene blue and proflavine are fluorescent dyes used to stain nucleic acid from the molecular level to the tissue level. Already clinically used for sentinel node mapping, detection of neuroendocrine tumors, methemoglobinemia, septic shock, ifosfamide-induced encephalopathy, and photodynamic inactivation of RNA viruses, the antimicrobial, anti-inflammatory, and antioxidant effect of methylene blue has been demonstrated in different in vitro and in vivo studies. Proflavine was used as a disinfectant and bacteriostatic agent against many gram-positive bacteria, as well as a urinary antiseptic involved in highlighting cell nuclei. At the tissue level, the anti-inflammatory effects of methylene blue protect against pulmonary, renal, cardiac, pancreatic, ischemic-reperfusion lesions, and fevers. First used for their antiseptic and antiviral activity, respectively, methylene blue and proflavine turned out to be excellent dyes for diagnostic and treatment purposes. In vitro and in vivo studies demonstrated that both dyes are efficient as perfusion and tissue tracers and permitted to evaluate the minimal efficient concentration in different species, as well as their pharmacokinetics and toxicity. This review aims to identify the optimal concentrations of methylene blue and proflavine that can be used for in vivo experiments to highlight the vascularization of the skin in the case of a perforasome (both as a tissue tracer and in vascular mapping), as well as their effects on tissues. This review is intended to be a comparative and critical presentation of the possible applications of methylene blue (MB) and proflavine (PRO) in the surgical field, and the relevant biomedical findings from specialized literature to date are discussed as well.

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“…7A-D). This analysis can be extremely effective in terms of ligand binding and the local dynamics associated with the binding [51]. The Figure here depicts an ensemble average of the top docking conformations.…”

Section: Ligand Induced Dissociation Of Ifabp Clusters As Probed By Imentioning

confidence: 99%

The Role of Intestinal Fatty Acid Binding Proteins in Protecting Cells from Fatty Acid Induced Impairment of Mitochondrial Dynamics and Apoptosis

Sarkar-Banerjee

Chowdhury

Sanyal

et al. 2018

Cell Physiol Biochem

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Background/Aims: The conformation, folding and lipid binding properties of the intestinal fatty acid binding proteins (IFABP) have been extensively investigated. In contrast, the functional aspects of these proteins are not understood and matter of debates. In this study, we aim to address the deleterious effects of FA overload on cellular components, particularly mitochondria; and how IFABP helps in combating this stress by restoring the mitochondrial dynamics. Methods: In the present study the functional aspect of IFABP under conditions of lipid stress was studied by a string of extensive in-cell studies; flow cytometry by fluorescence-activated cell sorting (FACS), confocal imaging, western blotting and quantitative real time PCR. We deployed ectopic expression of IFABP in rescuing cells under the condition of lipid stress. Again in order to unveil the mechanistic insights of functional traits, we arrayed extensive computational approaches by means of studying centrality calculations along with protein-protein association and ligand induced cluster dissociation. While addressing its functional importance, we used FCS and in-silico computational analyses, to show the structural distribution and the underlying mechanism of IFABP’s action. Results: Ectopic expression of IFABP in HeLa cells has been found to rescue mitochondrial morphological dynamics and restore membrane potential, partially preventing apoptotic damage induced by the increased FAs. These findings have been further validated in the functionally relevant intestinal Caco-2 cells, where the native expression of IFABP protects mitochondrial morphology from abrogation induced by FA overload. However, this native level expression is insufficient to protect against apoptotic cell death, which is rescued, at least partially in cells overexpressing IFABP. In addition, shRNA mediated IFABP knockdown in Caco-2 cells compromises mitochondrial dynamics and switches on intrinsic apoptotic pathways under FA-induced metabolic stress. Conclusion: To summarize, the present study implicates functional significance of IFABP in controlling ligand-induced damage in mitochondrial dynamics and apoptosis.

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Comparative Study of Toluidine Blue O and Methylene Blue Binding to Lysozyme and Their Inhibitory Effects on Protein Aggregation

Cited by 50 publications

References 77 publications

Lysozyme crystals dyed with bromophenol blue: where has the dye gone?

Lysozyme crystals dyed with bromophenol blue: where has the dye gone?

Comparative Study Regarding the Properties of Methylene Blue and Proflavine and Their Optimal Concentrations for In Vitro and In Vivo Applications

The Role of Intestinal Fatty Acid Binding Proteins in Protecting Cells from Fatty Acid Induced Impairment of Mitochondrial Dynamics and Apoptosis

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