Philo J S
Protein Chemistry Department, Amgen Inc., Thousand Oaks, California 91320, USA.
Biophys J. 1997 Jan;72(1):435-44. doi: 10.1016/S0006-3495(97)78684-3.
Many traditional approaches to the analysis of sedimentation velocity data work poorly with data for low-molecular-weight solutes, which have sedimentation boundaries that are severely broadened by diffusion. An approach that has previously had some success is to directly fit these broad boundaries to approximate solutions of the Lamm equation that directly account for the high diffusion. However, none of the available approximate solutions work well at times both early and late in the run, or give boundary shapes that are highly accurate, especially for species of molecular weight < 10,000. An improved fitting function has been developed to overcome some of these limitations. The new function adds two correction terms to the Fujita-MacCosham solution. The optimum coefficients for these new correction terms were determined by a least-squares approach. The accuracy and limitations of fitting with this new function were tested against synthetic data sets obtained by finite-element methods, for analysis of samples containing either single species or several noninteracting species. We also compare the strengths and weaknesses of this method of analysis, and its ability to work with noisy data, relative to recently developed time-derivative methodologies.
许多传统的沉降速度数据分析方法在处理低分子量溶质的数据时效果不佳,因为这些溶质的沉降边界会因扩散而严重展宽。一种此前取得了一定成功的方法是将这些宽边界直接拟合到Lamm方程的近似解,该近似解直接考虑了高扩散情况。然而,现有的近似解在实验前期和后期都不能很好地发挥作用,或者给出的边界形状不够精确,特别是对于分子量<10,000的物种。为克服其中一些局限性,已开发出一种改进的拟合函数。新函数在藤田-麦科沙姆解中添加了两个校正项。这些新校正项的最佳系数通过最小二乘法确定。针对通过有限元方法获得的合成数据集,测试了使用此新函数进行拟合的准确性和局限性,用于分析包含单一物种或几种非相互作用物种的样品。我们还比较了这种分析方法的优缺点,以及它相对于最近开发的时间导数方法处理噪声数据的能力。