20.500.12556/RUL-125374 The role of buffers in wild-type HEWL amyloid fibril formation mechanism Amyloid fibrils, highly ordered protein aggregates, play an important role in the onset of several neurological disorders. Many studies have assessed amyloid fibril formation under specific solution conditions, but they all lack an important phenomena in biological solutions—buffer specific effects. We have focused on the formation of hen egg-white lysozyme (HEWL) fibrils in aqueous solutions of different buffers in both acidic and basic pH range. By means of UV-Vis spectroscopy, fluorescence measurements and CD spectroscopy, we have managed to show that fibrillization of HEWL is affected by buffer identity (glycine, TRIS, phosphate, KCl-HCl, cacodylate, HEPES, acetate), solution pH, sample incubation (agitated vs. static) and added excipients (NaCl and PEG). HEWL only forms amyloid fibrils at pH = 2.0 under agitated conditions in glycine and KCl-HCl buffers of high enough ionic strength. Phosphate buffer on the other hand stabilizes the HEWL molecules. Similar stabilization effect was achieved by addition of PEG12000 molecules to the solution. lysozyme amyloid fibrils buffer-specific effects lizocim amiloidne fibrile pufer-specifični efekti true false true Angleški jezik Slovenski jezik Članek v reviji 2021-03-12 13:23:34 2021-03-12 13:23:35 2022-09-05 03:50:28 0000-00-00 00:00:00 2019 0 0 18 str. iss. 2, art. 65 Vol. 9 Feb. 2019 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.322 2218-273X 10.3390/biom9020065 1538128835 519952921 RAZ_Brudar_Sandi_2019.pdf RAZ_Brudar_Sandi_2019.pdf 1 56145EB7CC9BDCE1C7433165D0F062EF ecf348e509c697b0ce2d468300d8fdd06556d0cd6931b38241085775c3936538 b2ef1886-a1bb-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=141076 https://www.mdpi.com/2218-273X/9/2/65 1 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=141223 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0