Oxidative damage to β_L-crystallin in vitro by iron compounds formed in physiological buffers

β_L-crystallin aggregation due to oxidative damage in the presence of H₂O₂ and ferric chloride was studied in-vitro under conditions close to physiological. It was shown that the protein aggregation characterized by the nucleation time and the aggregation rate significantly depended on the composition of the isoosmotic buffers used, and decreased in the series HEPES buffer > Tris buffer > PBS. Ferric chloride at neutral pH was converted into water-insoluble iron hydroxide III (≡Fe^IIIOH). According to the data of scanning electron microscopy the ≡Fe^IIIOH particles formed in HEPES buffer, Tris buffer, and PBS practically did not differ in structure. However, the sizes of ≡Fe^IIIOH floating particles measured by dynamic light scattering differed significantly and were 44 ± 28 nm, 93 ± 66 nm, 433 ± 316 nm (Z_aver ± SD) for HEPES buffer, Tris buffer, and PBS, respectively. It was found by the spin trap method that the ability of ≡Fe^IIIOH to decompose H₂O₂ with the formation of a ^•OH decreases in the series HEPES buffer, Tris buffer, and PBS. The authors suggest that the ability to generate ^•OH during the decomposition of H₂O₂ is determined by the total surface area of ≡Fe^IIIOH particles, which significantly depends on the composition of the buffer in which these particles are formed.