Drug–protein binding: a critical review of analytical tools

Vuignier, Karine; Schappler, Julie; Veuthey, Jean‐Luc; Carrupt, Pierre‐Alain; Martel, Sophie

doi:10.1007/s00216-010-3737-1

Cited by 353 publications

(318 citation statements)

References 106 publications

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“…The binding of MO to HSA is the first stage of the color-change mechanism of the MO-HSA complex. The most suitable method for confirming the binding is an analysis performed using an SPR biosensor, which is an established method for verifying molecular interactions (5,10,15). The binding ratio of MO molecules to a single HSA molecule was analyzed by means of SPR analysis.…”

Section: Methodsmentioning

confidence: 99%

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Structure of the methyl orange-binding site on human serum albumin and its color-change mechanism

Ito

Yamamoto

2015

Biomed. Res.

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The goal in this study was to clarify the color-change mechanisms of methyl orange (MO) bound to human serum albumin (HSA) and the structure of the binding site. The absorbance of the MO-HSA complex was measured at 560 nm in solutions of varying pH (pH 2.4-6.6). The obtained pHdependent experimental data were consistent with the data calculated using the HendersonHasselbalch equation and pKa values (3.8, MO; 1.4, carboxyl group). The extent of the binding of MO to an HSA molecule was determined to be 1-4 by performing surface plasmon resonance analysis. Furthermore, the binding of MO to HSA was inhibited by warfarin. A fitting model of MO to HSA was created to evaluate these results based on PDB data (warfarin-HSA complex: 2BXD) and protein-structure analysis. The color-change mechanism of the MO-HSA complex appears to be as follows: the dissociated sulfo group of MO binds to Arg218/Lys444 sidechains through electrostatic interaction in the warfarin-binding site, and, subsequently, the color change occurs through a proton exchange between the diazenyl group and the γ-carboxyl group of Glu292. The color-changed MO is fixed in the warfarin-binding site. These results could support the development of a reliable dye-binding method and of a new method for staining diverse tissues that is based on a validated mechanism.Metachromasy is the characteristic color change of an acidic or basic dye used for staining biological tissues or cells (1, 7). The mechanism underlying metachromasy is considered to involve hydrophobic and hydrophilic interactions between basic (cationic) dyes, such as toluidine blue and methylene blue, and polyanions, such as nucleic acids and mucopolysaccharides containing phosphate and sulfate groups, respectively, and the resulting color change occurs through dye aggregation. In the case of acidic (anionic) dyes, the dyes interact with the amino groups of proteins (14, 16). Many pH indicators such as bromocresol green (BCG), bromocresol purple (BCP), and phenolphthalein exhibit metachromasy, and when albumin, which can adsorb various organic compounds (2), is added to a pH indicator in a buffered solution, the resulting color change in the indicator is equivalent to that which occurs in a solution of high or low pH. In clinical environments, dye-binding methods involving the use of BCG and BCP that are employed for quantifying human serum albumin (HSA) are based on this property of albumin (3). Because the color change occurs in a buffered solution whose pH is constant, a proton-exchange reaction occurs between the molecules. In the case of pH indicators, the amount of pH-dependent color change is measured using a photometric method after the binding of the indicator to HSA, and the amount is calculated using the Henderson-Hasselbalch equation. In our previous work, a comparison between experimental and calculated pH-dependent data indicated that color change

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Section: Methodsmentioning

confidence: 99%

“…Binding was analyzed by performing a steady-state affinity analysis by using the BIAevaluation program. After completion of the assay procedure, the surface was regenerated using 50 mmol/L NaOH (5,10,15).…”

mentioning

confidence: 99%

Structure of the methyl orange-binding site on human serum albumin and its color-change mechanism

Ito

Yamamoto

2015

Biomed. Res.

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“…1 The protein binding of drugs is an important pharmacokinetic parameter affecting their biological activity, metabolism, distribution, and elimination. 2,3 The magnitude of the effect of a drug on the target organ is related to its free concentration. Human serum albumin (HSA) is the most abundant protein with a molecular weight of 65 -69 KDa.…”

Section: Introductionmentioning

confidence: 99%

Interactions Between Sirolimus and Anti-Inflammatory Drugs: Competitive Binding for Human Serum Albumin

Khodaei¹,

Bolandnazar²,

Valizadeh³

et al. 2016

Adv Pharm Bull

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“…Consequently, the determination of affinity constants that describe the interaction strengths between drugs and plasma proteins is very important during the drug discovery phase when screening NCE. Several approaches have been used to measure drug-protein interactions, such as equilibrium dialysis, calorimetric and spectroscopic methods, affinity cluomatography, and biosensor-based assays [ 62]. These methods nevertheless present the same drawbacks mentioned for pKa and log P determination, in addition to specific ones including protein and/or drug adsorption and immobilization of the protein onto a support, which can alter the binding properties of the protein of interest.…”

Section: Plasma Protein Bindingmentioning

confidence: 99%