Spectroscopic analysis to identify the binding site for Rifampicin on Bovine Serum Albumin

Sharma, Sudhanshu; Takkella, Dineshbabu; Kumar, Pintu; Gavvala, Krishna

doi:10.1016/j.saa.2022.121721

Cited by 7 publications

(7 citation statements)

References 71 publications

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“…The addition of protein BSA resulted in a strong increase in TB-p and TB-AA emission ( Figure 2 b), with titration data fitting nicely to the 1:1 stoichiometry of the dye/BSA complex. Thus, compounds bind to the BSA with biologically relevant 10 µM affinity ( Table 1 ), likely in one of the binding sites for small molecules [ 17 ].…”

Section: Resultsmentioning

confidence: 99%

Triarylborane-“Click” Fluorescent Tag for Orthogonal Amino Acid Labelling, Interactions with DNA, Protein, and Cyclodextrins

Jurković,

Ferger,

Drašković

et al. 2023

Pharmaceuticals

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The innovative design of a triarylborane (TB)-dye with one NMe2-alkylated (propargylated) group and one NMe2 group yielded a system that is both an NMe2 π-donor and an inductive NMe2-alkyl cationic acceptor. Consequently, the new TB-dye was highly sensitive to a “click” reaction with an azide-substituted lysine side chain (yielding TB-lysine), resulting in a bathochromic shift of emission of 100 nm. In addition, fluorene attached to the lysine C-terminus showed FRET with the TB-chromophore, also sensitive to interactions with targets. Both the TB-dye and TB-lysine showed high affinities towards both DNA and proteins, reporting binding by an opposite fluorimetric response for DNA/RNA (quenching) vs. BSA (increase). Thus, the novel TB-dye is an ideal fluorimetric probe for orthogonal incorporation into bio-targets by “click” reactions due to fluorescence reporting of the progress of the “click” reaction and further sensing of the binding site composition. The TB-dye is moderately toxic to human cell lines after 2–3 days of exposure, but efficiently enters cells in 90 min, being non-toxic at short exposure. The most important product of the “click” reaction, TB-lysine, was non-toxic to cells and showed equal distribution between mitochondria and lysosomes. Further studies would focus particularly on the very convenient monitoring of the progress of “click” conjugation of the TB-dye with biorelevant targets inside living cells by confocal microscopy.

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Section: Resultsmentioning

confidence: 99%

Triarylborane-“Click” Fluorescent Tag for Orthogonal Amino Acid Labelling, Interactions with DNA, Protein, and Cyclodextrins

Jurković,

Ferger,

Drašković

et al. 2023

Pharmaceuticals

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“…The greater lifetime is attributed to the solvent-exposed residue or Trp-134, whereas the lower lifetime has been attributed to buried Trp or Trp-213. 42,62 The fluorescence lifetime in the presence of ANSA was observed at an emission wavelength of 470 nm. Figure 9 and Tables 7−9 represent the lifetime of BSA in the presence of ANSA at pH 5, 7.4, and 10.…”

Section: Time-correlated Single Photonmentioning

confidence: 99%

Analytical Aspects of ANSA–BSA Association: A Thermodynamic and Conformational Approach

Jain,

Judy,

Kishore

2024

J. Phys. Chem. B

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Many studies have demonstrated the manner in which ANS interacts with bovine serum albumin (BSA), although they are limited by the extremely low solubility of dye. The present study demonstrates the binding of ANSA dye with BSA, and since this dye can easily replace ANS, it not only simplifies research but also improves sensor accuracy for serum albumin. A combination of calorimetry and spectroscopy has been employed to establish the thermodynamic signatures associated with the interaction of ANSA with the protein and the consequent conformational changes in the latter. The results of differential scanning calorimetry reveal that when the concentration of ANSA in solution is increased, the thermal stability of the protein increases substantially. The fluorescence data demonstrated a decrease in the binding affinity of ANSA with the protein when pH increased but was unable to identify a change in the mode of interaction of the ligand. ITC has demonstrated that the mode of interaction between ANSA and the protein varies from a single set of binding sites at pH 5 and 7.4 to a sequential binding site at pH 10, emphasizing the potential relevance of protein conformational changes. TCSPC experiments suggested a dynamic type in the presence of ANSA. Molecular docking studies suggest that ANSA molecules are able to find ionic centers in the hydrophobic pockets of BSA. The findings further imply that given its ease of use in experiments, ANSA may be a useful probe for tracking the presence of serum albumin and partially folded protein states.

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“…The alarming data cited above is due to problems associated with the currently recommended therapeutic regimen, which consists of chemotherapy based on the administration of four antibiotics: isoniazid, rifampicin, pyrazinamide, and ethambutol [ 2 , 4 , 5 ]. These drugs are administered in high doses and for a prolonged period of time, which, depending on the patient’s response to treatment, can last up to 9–12 months, causing the appearance of serious side effects.…”

Section: Introductionmentioning

confidence: 99%

“…Another worrying factor is the development of bacterial resistance to these antibiotics, which can lead to the emergence of multidrug-resistant TB. This problem has been declared a global health emergency by the WHO and may jeopardize disease control worldwide [ 1 , 2 , 5 ].…”

Section: Introductionmentioning

confidence: 99%

Montmorillonite–Rifampicin Nanohybrid for pH-Responsive Release of the Tuberculostatic

et al. 2023

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The present work describes the development of a hybrid and pH-responsive system for rifampicin using the clay mineral ‘montmorillonite’ as a nanocarrier. The influence of operational variables on the drug incorporation process was evaluated using 24 factorial designs. Under optimized conditions, the experiment allowed an incorporated drug dose equivalent to 98.60 ± 1.21 mg/g. Hybrid systems were characterized by different characterization techniques (FTIR, XRD, TGA, DSC, and SEM) to elucidate the mechanism of interaction between the compounds used. Through in vitro release studies, it was possible to verify the efficacy of the pH-dependent system obtained, with approximately 70% of the drug released after sixteen hours in simulated intestinal fluid. The adjustment of the experimental release data to the theoretical model of Higuchi and Korsmeyer–Peppas indicated that the release of rifampicin occurs in a prolonged form from montmorillonite. Elucidation of the interactions between the drug and this raw clay reinforces its viability as a novel carrier to develop an anti-TB/clay hybrid system with good physical and chemical stability.

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Spectroscopic analysis to identify the binding site for Rifampicin on Bovine Serum Albumin

Cited by 7 publications

References 71 publications

Triarylborane-“Click” Fluorescent Tag for Orthogonal Amino Acid Labelling, Interactions with DNA, Protein, and Cyclodextrins

Triarylborane-“Click” Fluorescent Tag for Orthogonal Amino Acid Labelling, Interactions with DNA, Protein, and Cyclodextrins

Analytical Aspects of ANSA–BSA Association: A Thermodynamic and Conformational Approach

Montmorillonite–Rifampicin Nanohybrid for pH-Responsive Release of the Tuberculostatic

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