Prokhorov General Physics Institute of the Russian Academy of Sciences (GPI RAS), 119991 Moscow, Russia.
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997 Moscow, Russia.
Biomolecules. 2022 Nov 1;12(11):1613. doi: 10.3390/biom12111613.
The properties of a lysozyme solution under laser-induced breakdown were studied. An optical breakdown under laser action in protein solutions proceeds with high efficiency: the formation of plasma and acoustic oscillations is observed. The concentration of protein molecules has very little effect on the physicochemical characteristics of optical breakdown. After exposure to optical breakdown, changes were observed in the enzymatic activity of lysozyme, absorption and fluorescence spectra, viscosity, and the sizes of molecules and aggregates of lysozyme measured by dynamic light scattering. However, the refractive index of the solution and the Raman spectrum did not change. The appearance of a new fluorescence peak was observed upon excitation at 350 nm and emission at 434 nm at exposure for 30 min. Previously, a peak in this range was associated with the fluorescence of amyloid fibrils. However, neither the ThT assay nor the circular dichroism dispersion confirmed the formation of amyloid fibrils. Probably, under the influence of optical breakdown, a small part of the protein degraded, and a part changed its native state and aggregated, forming functional dimers or "native aggregates".
研究了溶菌酶溶液在激光诱导击穿下的性质。在蛋白质溶液中,激光作用下的光学击穿以高效率进行:观察到等离子体和声波振荡的形成。蛋白质分子的浓度对光学击穿的物理化学特性几乎没有影响。在经受光学击穿后,观察到溶菌酶的酶活性、吸收和荧光光谱、粘度以及通过动态光散射测量的溶菌酶分子和聚集体的大小发生了变化。然而,溶液的折射率和拉曼光谱没有变化。在暴露 30 分钟后,在 350nm 激发和 434nm 发射时观察到新的荧光峰的出现。以前,该范围内的峰与淀粉样纤维的荧光有关。然而,ThT 测定和圆二色性色散都没有证实淀粉样纤维的形成。可能是在光学击穿的影响下,一部分蛋白质降解,一部分蛋白质改变其天然状态并聚集,形成功能性二聚体或“天然聚集体”。