Parameters Native albumin Nitroxidized-albumin (0.75 mMperoxynitrite) Nitroxidized albumin (1.5 mMperoxynitrite) Changes on modification Absorbance at 278 nm 0.518 0.727 0.849 29 and 39% hyperchromicity Absorbance at 428 nm ---- 0.204 0.291 ---- Nitrotyrosine content (nmole mg-1 HSA) ---- 48.5 69.2 ---- Nitrotryptophan content (nmole mg-1 HSA) ---- 33.6 52.6 ---- aFluorescence intensity, (F.I.) bλexc = 275 nm 163 43.3 29.9 73 and 81% decrease in F.I. bλexc = 295 nm 51.7 15.9 10.9 69 and 79% decrease in F.I. Protein carbonyl content (nmole mg-1 HSA) 2.5 21.3c 30.3c 88 and 92% increase Dityrosine content (nmole mg-1 HSA) ---- 25.4 52.2 ---- Dityrosine aF.I.  bλexc = 325 nm ---- 22.7 95.7 ---- Far-UV CD  dMRE at 208 nm -21264.1 -18367.7 -14189.7 ---- dMRE at 222 nm -20057.4 -16493.7 -11733.1 ---- dMRE at 192 nm 42517.1 38063.5 24200.6 ---- α-helix (%) 68.28 60.4 40.1 ---- ThT binding, aF.I.  bλexc = 435 nm 33.9 91.7 158.5 63 and 77% increase in F.I. aFluorescence intensity (F.I.) in arbitrary units (A.U.) bλexc = excitation wavelength cp<0.0001 vs. native albumin dMRE value in deg cm2 dmol-1

Table 1: Physicochemical characteristic of native and nitroxidized-albumin.