Characterization of apoA-I variants’ conformation.

<p>A) Intrinsic Trp fluorescence emission spectra of apoA-I variants. Proteins at a final concentration of 0.1 mg/mL in citrate phosphate Mc Ilvaines buffer pH 7.4. Excitation was set at 295 nm and emission was recorded between 310 and 370 nm. Continuous line represents Wt protein; Dashed and dotted lines are fluorescence spectra corresponding to Gly26Arg and Lys107-0 respectively. B) Fluorescence analysis of bis-ANS binding to apoA-I. ApoA-I variants, at a final concentration of 0.1 mg/mL were titrated with bis-ANS to a final concentration of 16 µM. The probe was excited at 360 nm, and emission registered as the Wavelength of Maximum Fluorescence for this probe. Dark circles represent the experimental data for Wt. Gray and white symbols correspond to Lys107-0 and Gly26Arg respectively. Inset: normalized fluorescence spectra of bis-ANS at a molar ratio 1∶1 probe to protein. Continuous line corresponds to Wt spectrum; Dashed and dotted lines correspond to Lys107-0 and Gly26Arg respectively.