Ligand-induced thermostability in proteins: Thermodynamic analysis of ANS–albumin interaction

Celej, M. Soledad; Dassie, Sergio Alberto; Freire, Eleonora; Bianconi, M. Lucia; Fidelio, Gerardo D.

doi:10.1016/j.bbapap.2005.05.003

Cited by 57 publications

(55 citation statements)

References 48 publications

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“…32,33) In this sense, we have reported that aniline naphthalene sulfonate (ANS) derivatives induce an overall tightening of the BSA structure which may contribute to an unfavorable conformational entropy change. 24,34) The positive entropy contribution observed for AHTC-BSA interaction suggests the release of structural water molecules involved in a network of interactions. 32,33) DSC Analysis Protein stability is modulated by ligands that bind with a moderate or high affinity as a thermodynamic consequence of the coupling of the unfolding and binding equilibria.…”

Section: Fluorescence Changes Upon Ahtc-bsa Interactionmentioning

confidence: 99%

“…This effect is accounted for by a shift of T m along with a change in DH m , being more marked for high affinity protein-ligand interactions. 24,26,35,36) We employed DSC as an analytical tool to estimate the binding parameters. Figure 5 shows representative experimental endotherms obtained for BSA alone and at increasing [AHTC] tot /[BSA] tot before (inset) and after baseline subtraction.…”

Section: Fluorescence Changes Upon Ahtc-bsa Interactionmentioning

confidence: 99%

“…The protein unfolding parameters were in excellent agreement with those reported previously. 24) The cooperative unit was near 1 (Table 2) suggesting a two-state unfolding process 22) (see DSC measurements section in Materials and Methods).…”

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confidence: 99%

“…35,36) This dependence allows the estimation of n if model-based expressions for the binding polynomial are supplied. 36 24,36) : (2) where R is the gas constant, A is a constant, and the other parameters are as defined previously. We have successfully used this approach to estimate the number of binding sites for different dyes on serum albumins.…”

mentioning

confidence: 99%

“…23) Reversible unfolding models were employed to fit the data after checking that the calorimetric signal was reduced by about 40% after heating a BSA solution up to the T m , then cooled it to room temperature and then reheated it to TӷT m and that similar thermograms and results were obtained either (i) at different protein concentrations (ref. 24 and this work) and (ii) by fitting the entire DSC curve using only the first half of the experimental data from the endotherm.…”

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Binding of the Highly Toxic Tetracycline Derivative, Anhydrotetracycline, to Bovine Serum Albumin

Burgos

Fernández

Celej

et al. 2011

Biological & Pharmaceutical Bulletin

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Tetracycline (TC) derivatives comprise a group of bacteriostatic antibiotics extensively used in human and veterinary medicine and in acquaculture, and as animal growth promoters due to their safe toxicological profile.1) More recently, they have been proposed as anti-amyloidogenic drugs since they can interfere with protein conformational changes and self-assemblies associated with protein misfolding and deposition diseases. 2-4)Despite their popular use, TC derivatives are highly unstable and some degradation products formed during the storage or normal use of the drugs are pharmacologically toxic or inactive compounds.5) Anhydrotetracycline (AHTC) is one of the major degradation products of TC that has been linked to several side effects of the drug, e.g., cutaneous phototoxicity 6) and Fanconi-type syndrome. 7) AHTC has also proven to be more toxic than TC against environmentally relevant bacteria. 8)AHTC can be produced by dehydration in acid media, 5,8) by photolysis in mild reducing conditions 6,9,10) or during high-temperature treatments.11) Removal of a water molecule from the C5a-C6 positions of TC by treatment with warm mineral acid gives the anhydroderivative (Fig. 1).12) Dehydration leads to significant changes in aromatization and planarity of the tetracyclic moiety, 13,14) showing variations in potency, 15) mechanism of action 1) and lipophilicity 16,17) as compared to the parent compound. Changes in chemical configuration can also modulate the interaction of drugs with serum proteins. 18)Serum albumins are the main carriers of physiological metabolites and pharmacological drugs in blood. Protein binding has a significant impact on the pharmacokinetics and biological activity of drugs and modulates the toxicity of bound substances, attenuates adverse reactions and prolongs the in vivo half-life of therapeutic agents. 18-21)The facts outlined above prompted us to investigate the interaction of AHTC with bovine serum albumin (BSA) using three biophysical techniques. Binding affinity and number of interacting ligands were assessed by fluorimetric titration. . Drug binding was enthalpically and entropically driven and seemed to involve hydrophobic interactions. AHTC fluorescence enhancement and hypsochromic shifts observed upon binding suggested a low-polarity location excluded from water for the bound drug. Our data are useful for evaluating the biodisponibility of the pharmacophore and the dynamic distribution of the toxic derivative.

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Section: Fluorescence Changes Upon Ahtc-bsa Interactionmentioning

confidence: 99%

Section: Fluorescence Changes Upon Ahtc-bsa Interactionmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Binding of the Highly Toxic Tetracycline Derivative, Anhydrotetracycline, to Bovine Serum Albumin

Burgos

Fernández

Celej

et al. 2011

Biological & Pharmaceutical Bulletin

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Molecular cloning, expression profile, odorant affinity, and stability of two odorant‐binding proteins in Macrocentrus cingulum Brischke (Hymenoptera: Braconidae)

Ahmed

Zhang

Wang

et al. 2017

Arch Insect Biochem Physiol

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The polyembryonic endoparasitoid wasp Macrocentrus cingulum Brischke (Hymenoptera: Braconidae) is deployed successfully as a biocontrol agent for corn pest insects from the Lepidopteran genus Ostrinia in Europe and throughout Asia, including Japan, Korea, and China. The odorants are recognized, bound, and solubilized by odorant-binding protein (OBP) in the initial biochemical recognition steps in olfaction that transport them across the sensillum lymph to initiate behavioral response. In the present study, we examine the odorant-binding effects on thermal stability of McinOBP2, McinOBP3, and their mutant form that lacks the third disulfide bonds. Real-time PCR experiments indicate that these two are expressed mainly in adult antennae, with expression levels differing by sex. Odorant-binding affinities of aldehydes, terpenoids, and aliphatic alcohols were measured with circular dichroism spectroscopy based on changes in the thermal stability of the proteins upon their affinities to odorants. The obtained results reveal higher affinity of trans-caryophelle, farnesene, and cis-3-Hexen-1-ol exhibits to both wild and mutant McinOBP2 and McinOBP3. Although conformational flexibility of the mutants and shape of binding cavity make differences in odorant affinity between the wild-type and mutant, it suggested that lacking the third disulfide bond in mutant proteins may have chance to incorrect folded structures that reduced the affinity to these odorants. In addition, CD spectra clearly indicate proteins enriched with α-helical content.

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A thermodynamic characterization of the interaction of 8‐anilino‐1‐naphthalenesulfonic acid with native globular proteins: the effect of the ligand dimerization in the analysis of the binding isotherms

Andújar‐Sánchez

Jara‐Pérez

Cobos

et al. 2011

J of Molecular Recognition

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8-Anilino-1-naphthalenesulfonic acid (ANS) is a popular fluorescence probe, broadly used for the analysis of proteins, but the nature of its interaction with proteins and the high increase in the fluorescence intensity that takes place upon such process are still unclear. In the last few years, isothermal titration calorimetry has been used to characterize the nature of the interaction of this dye with proteins. The analysis of the binding isotherms of these studies has not considered the dimerization equilibrium of ANS, which is pH dependent, and it can result in serious errors in the data analysis. In the present work we have developed a suitable data analysis by which this process is taken into account. To study the binding of the dye to proteins at different pH values, we have used the Abl-SH3 domain. Our results suggest that at pH 3 and 5, where the dimerization of the ANS is important, electrostatic interactions are significant for the binding of ANS to the Abl-SH3 domain. However, at pH 7, ANS behaves mostly as monomer and the interaction with the protein is mainly hydrophobic. The pH dependent behavior of the ANS binding to proteins can be explained in terms of ionization states of both, the protein and the ANS.

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Ligand-induced thermostability in proteins: Thermodynamic analysis of ANS–albumin interaction

Cited by 57 publications

References 48 publications

Binding of the Highly Toxic Tetracycline Derivative, Anhydrotetracycline, to Bovine Serum Albumin

Binding of the Highly Toxic Tetracycline Derivative, Anhydrotetracycline, to Bovine Serum Albumin

Molecular cloning, expression profile, odorant affinity, and stability of two odorant‐binding proteins in Macrocentrus cingulum Brischke (Hymenoptera: Braconidae)

A thermodynamic characterization of the interaction of 8‐anilino‐1‐naphthalenesulfonic acid with native globular proteins: the effect of the ligand dimerization in the analysis of the binding isotherms

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