Measurement of biomolecular interactions and association via light scattering

通过光散射测量生物分子相互作用和缔合

• 批准号: 8939504
• 负责人: Allen P Minton
• 金额: $ 12.67万
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8939504
• 关键词: Charge; Dependence; Diffusion; Fluorescence; Laboratories; Light; Measurement; Measures; Modeling; Molecular Conformation; Monitor; Ovalbumin; Ovomucin; Pancreatic ribonuclease; Proteins; Reproducibility; Ribonucleases; Techniques; Thermodynamics; Ursidae Family; Work; light scattering; macromolecule; particle; research study

1. The concentration dependence of light scattering of three proteins, ovalbumin, ribonuclease A, and ovomucoid, and binary mixtures of the three proteins were measured at various compositions and pH values at total protein concentrations of up to 35 g/l, under conditions such that ribonuclease bears a net positive charge and the other proteins bear a net negative charge. The concentration dependence was interpreted in the context of three models: a statistical thermodynamic model and two extensions of the effective hard particle model generalized to allow for the presence of non-additive interactions between species in a binary mixture (D. Wu, A. Minton). 2. Experiments were performed to determine whether quantitative composition gradient light scattering experiments can be performed using a fluorescence spectrophotometer in place of a dedicated light scattering photometer. Results obtained so far are encouraging, but further work will be required in order to achieve the requisite reproducibility.

1.测定了卵清蛋白、核糖核酸酶A和卵粘蛋白三种蛋白质及其二元混合物在不同组成和pH值下的光散射随浓度的变化关系。在总蛋白质浓度高达35g/L时，核糖核酸酶带净正电荷，其余蛋白质带净负电荷。浓度依赖性是在三个模型的背景下解释的：一个统计热力学模型和两个推广的有效硬粒子模型，以允许二元混合物中物种之间存在非加成相互作用(D.Wu，A.Minton)。 2.用荧光分光光度计代替专用的光散射光度计进行定量成分梯度光散射实验。迄今取得的成果令人鼓舞，但还需要进一步的工作，以实现必要的可重复性。