Multi-angle static and dynamic light scattering investigation of lysozyme association: From crystallization to liquid-liquid phase separation
Multi-angle static and dynamic light scattering investigation of lysozyme association: From crystallization to liquid-liquid phase separation
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DOI：
10.1016/j.jcrysgro.2007.03.009
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发表时间：
2007-06-15
影响因子：
1.8
通讯作者：
Kanzaki, Noriko
中科院分区：
文献类型：
--
作者：
Onuma, Kazuo;Kanzaki, Noriko
使用时间分辨多角度静态和动态光散射观察鸡蛋清溶菌酶的关联。在含有 1.2M NaCl、pH 为 4.6、用 50 mM NaAc 缓冲并维持在 20°C 的溶液中，使用静态光散射以 0.3-5 秒的间隔测量 8 至 32 mg/mL 的各种溶菌酶浓度的溶菌酶聚集体的表观分子量 M-w 和回转半径 R-g。在 32 mg/mL 时，发生液-液相分离 (LLPS)。 (Kc/Delta R(q))(1/2) 与 q(2) 图具有非线性形状，在特定 q(2) 处具有最大值，表明聚集体尺寸分布在较窄的范围内。在我们之前对高分子量蛋白质的研究中没有观察到这一点。动态光散射数据与静态光散射数据一致；即，CONTIN 分析显示出清晰的双峰（单体和聚集体）分布。对聚集体与蛋白质浓度的分形维数分析表明，LLPS 过程中形成的散射体的内部结构在最后阶段是刚性的，这支持两步成核模型。 (c) 2007 Elsevier B.V. 保留所有权利。
The association of hen egg white lysozyme was observed using time-resolved multi-angle static and dynamic light scattering. In solutions containing 1.2M of NaCl with a pH of 4.6 buffered by 50 mM of NaAc, and maintained at 20 degrees C, the apparent molecular weight M-w and gyration radius R-g, of lysozyme aggregates were measured at 0.3-5-s intervals for various lysozyme concentrations from 8 to 32 mg/mL using static light scattering. At 32 mg/mL, liquid-liquid phase separation (LLPS) occurred. The (Kc/Delta R(q))(1/2) vs. q(2) plot had a non-linear shape with a maximum at a particular q(2), indicating that the aggregate size distribution was within a narrow range. This had not been observed in our previous studies of high-molecular-weight proteins. The dynamic light scattering data were consistent with those of static light scattering; i.e., CONTIN analysis showed clear bimodal (monomer and aggregate) distributions. Fractal dimension analysis of the aggregates with the protein concentration showed that the inner structure of the scatterer formed during LLPS was rigid in the final stage, which supports a two-step nucleation model. (c) 2007 Elsevier B.V. All rights reserved.