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沉降系数和分子量的同步快速估算

Simultaneous rapid estimation of sedimentation coefficient and molecular weight.

作者信息

Holladay L A

机构信息

Department of Biochemistry, Vanderbilt University, Nashville, Tennessee 37232, USA.

出版信息

Biophys Chem. 1980 Apr;11(2):303-8. doi: 10.1016/0301-4622(80)80033-0.

Abstract

Data obtained from the early portion of sedimentation velocity experiments may be analyzed to simultaneously estimate both s and s/D. The C versus r data obtained are analyzed using a nonlinear least squares algorithm and an approximate solution to the Lamm equation. This procedure was tested both with simulated noisy data and with experimental data obtained using ribonuclease, ovalbumin, and somatostatin.dodecylsulfate. The procedure assumes that both s and D are independent of concentration. The results suggest that optimal estimation of both s and s/D is obtained at values of (= 2D/(s.omega(2).r(a)(2))) in the range of 0.002 to 0.01 and values of tau(= 2 somega(2)t) less than 0.04. Appropriate selection of rotor speed allows the estimation of both s and s/D for nearly globular macromolecules in the range of 10(4) to 10(6) daltons with data obtained during the first 3000-5000 seconds of a sedimentation velocity experiment.

摘要

从沉降速度实验早期获得的数据可用于分析,以同时估算沉降系数(s)和沉降扩散系数(s/D)。利用非线性最小二乘法算法和Lamm方程的近似解对获得的浓度(C)与径向距离(r)的数据进行分析。该程序在模拟噪声数据以及使用核糖核酸酶、卵清蛋白和生长抑素 - 十二烷基硫酸盐获得的实验数据上都进行了测试。该程序假定沉降系数(s)和扩散系数(D)均与浓度无关。结果表明,当(= 2D/(s.ω².rₐ²))的值在0.002至0.01范围内且(τ = 2ω²t)的值小于0.04时,可获得沉降系数(s)和沉降扩散系数(s/D)的最佳估算值。通过适当选择转子速度,利用沉降速度实验最初3000 - 5000秒内获得的数据,可对分子量在10⁴至10⁶道尔顿范围内的近似球状大分子的沉降系数(s)和沉降扩散系数(s/D)进行估算。

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