Effect of pH on HSA-induced chirality in dansylglycine.

<p>(<b>a</b>) HSA (30 µM) was incubated at various pHs. (<b>b</b>) CD spectra after addition of 30 µM dansylglycine. (<b>c</b>) Absorbance spectra after addition of 30 µM dansylglycine. (<b>d</b>) Absorbance spectra of 30 µM dansylglycine alone.</p

Induced ellipticity as a function of dansylglycine concentration.

<p>The mixtures consisted of 15 µM HSA and increasing concentration of DG, as indicated. The results are the average and SD of experiments performed in triplicate.</p

Temperature-dependent reversibility of ICD in dansylglycine.

<p>The mixtures consisted of 30 µM HSA and 30 µM DG. (<b>a</b>) The temperature was increased from 20°C to 45°C and (<b>b</b>) returned to 20°C at a rate of 0.5°C/min.</p

HSA-Induced ellipticity in dansylglycine.

<p>(<b>a</b>) Near-UV-CD and (<b>b</b>) UV-Vis spectra of dansylglycine in the presence or absence of HSA. The mixtures consisted of 30 µM HSA and 30 µM DG.</p

Temperature-induced denaturation of HSA and its effect on ICD of dansylglycine.

<p>(<b>a</b>) Denaturation curve of HSA. The temperature was increased at 1°C/min, equilibrated at the target temperature for 2 min before each measurement and reversed following the same protocol; (<b>b</b>) DG was added before denaturation and the ICD spectrum measured at 25°C; (<b>c</b>) DG was added after denaturation and the ICD spectrum measured at 25°C.</p

Quenching of the intrinsic fluorescence of HSA by dansylglycine and determination of binding constant.

<p>(<b>a</b>) Emission spectra of HSA (5 µM; λ_ex = 295 nm) in the presence of dansylglycine (DG; 0–35 µM). (<b>b</b>) Stern-Volmer plots at different temperatures. (<b>c</b>) Non-linear fitting for determination of the binding constant (Ka). <b>(Table)</b> Stern-Volmer and binding constants at different temperatures. The results are the average and SD of experiments performed in triplicate.</p

Displacement of dansylglycine (DG) from HSA by site I and II ligands: circular dichroism studies.

<p>The mixtures consisted of 30 µM HSA, 30 µM DG, and 30 µM of naproxen (NX), ibuprofen (IB), phenylbutazone (PB), or warfarin (WF).</p

Effect of oxidation on HSA-induced chirality.

<p>The mixtures consisted of 30 µM HSA and (<b>a</b>) 600 µM or (<b>b</b>) 1200 µM of the oxidants. Dansylglycine (30 µM) was added after oxidation.</p