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Molecular mass determination by sedimentation velocity experiments and direct fitting of the concentration profiles.

机译:通过沉降速度实验和浓度分布的直接拟合确定分子量。

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摘要

A new method for the direct molecular mass determination from sedimentation velocity experiments is presented. It is based on a nonlinear least squares fitting procedure of the concentration profiles and simultaneous estimation of the sedimentation and diffusion coefficients using approximate solutions of the Lamm equation. A computer program, LAMM, was written by using five different model functions derived by Fujita (1962, 1975) to describe the sedimentation of macromolecules during centrifugation. To compare the usefulness of these equations for the analysis of hydrodynamic results, the approach was tested on data sets of Claverie simulations as well as experimental curves of some proteins. A modification for one of the model functions is suggested, leading to more reliable sedimentation and diffusion coefficients estimated by the fitting procedure. The method seems useful for the rapid molecular mass determination of proteins larger than 10 kDa. One of the equations of the Archibald type is also suitable for compounds of low molecular mass, probably less than 10 kDa, because this model function requires neither the plateau region nor a meniscus free of solute.
机译:提出了一种通过沉降速度实验直接测定分子量的新方法。它基于浓度分布的非线性最小二乘拟合程序,并使用Lamm方程的近似解同时估计沉降和扩散系数。使用藤田(1962,1975)导出的五个不同的模型函数编写了一个计算机程序LAMM,以描述离心过程中大分子的沉积。为了比较这些方程式对水动力结果分析的有用性,在Claverie模拟的数据集以及某些蛋白质的实验曲线上对该方法进行了测试。建议对模型功能之一进行修改,以使拟合程序估算出更可靠的沉降和扩散系数。该方法似乎对快速测定分子量大于10 kDa的蛋白质很有用。 Archibald类型的方程式之一也适用于分子量可能小于10 kDa的低分子量化合物,因为该模型函数既不需要平稳区域也不需要弯液面,而无需溶质。

著录项

  • 作者

    Behlke, J; Ristau, O;

  • 作者单位
  • 年度 1997
  • 总页数
  • 原文格式 PDF
  • 正文语种 en
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