Quality control of protein standards for molecular mass determinations by small-angle X-ray scattering
Quality control of protein standards for molecular mass determinations by small-angle X-ray scattering
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DOI：
10.1107/s002188981000138x
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发表时间：
2010-04-01
影响因子：
6.1
通讯作者：
Akiyama, Shuji
中科院分区：
文献类型：
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作者：
Akiyama, Shuji
小角 X 射线散射 (SAXS) 是一种强大的技术，可用于评估溶液中生物大分子的尺寸和形状。相对于粒子浓度归一化的前向散射强度 I(0)/c 可用作分子质量的良好测量。从 SAXS 数据推导分子量的一般方法是确定目标蛋白与已知分子量的标准蛋白的 I(0)/c 之比。这种蛋白质间校准的准确性很大程度上受到所制备标准品的单分散性以及估计其浓度的精度的影响。在本研究中，提出了色谱分级分离和流体动力学表征作为制备一系列单分散蛋白质标准品的有效程序。使用单分散牛血清白蛋白作为标准，证明了分子量估计的平均偏差在 8% 之内。目前的结果证明了蛋白质标准质量控制的重要性，以充分利用蛋白质间校准的优势。
Small-angle X-ray scattering (SAXS) is a powerful technique with which to evaluate the size and shape of biological macromolecules in solution. Forward scattering intensity normalized relative to the particle concentration, I(0)/c, is useful as a good measure of molecular mass. A general method for deducing the molecular mass from SAXS data is to determine the ratio of I(0)/c of a target protein to that of a standard protein with known molecular mass. The accuracy of this interprotein calibration is affected considerably by the monodispersity of the prepared standard, as well as by the precision in estimating its concentration. In the present study, chromatographic fractionation followed by hydrodynamic characterization is proposed as an effective procedure by which to prepare a series of monodispersed protein standards. The estimation of molecular mass within an average deviation of 8% is demonstrated using monodispersed bovine serum albumin as a standard. The present results demonstrate the importance of protein standard quality control in order to take full advantage of interprotein calibration.