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小肽沉降系数的测定

Determination of sedimentation coefficients for small peptides.

作者信息

Schuck P, MacPhee C E, Howlett G J

机构信息

Biomedical Engineering and Instrumentation Program, ORS, National Institutes of Health, Bethesda, Maryland 20892, USA.

出版信息

Biophys J. 1998 Jan;74(1):466-74. doi: 10.1016/S0006-3495(98)77804-X.

DOI:10.1016/S0006-3495(98)77804-X
PMID:9449347
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1299399/
Abstract

Direct fitting of sedimentation velocity data with numerical solutions of the Lamm equations has been exploited to obtain sedimentation coefficients for single solutes under conditions where solvent and solution plateaus are either not available or are transient. The calculated evolution was initialized with the first experimental scan and nonlinear regression was employed to obtain best-fit values for the sedimentation and diffusion coefficients. General properties of the Lamm equations as data analysis tools were examined. This method was applied to study a set of small peptides containing amphipathic heptad repeats with the general structure Ac-YS-(AKEAAKE)nGAR-NH2, n = 2, 3, or 4. Sedimentation velocity analysis indicated single sedimenting species with sedimentation coefficients (s(20,w) values) of 0.37, 0.45, and 0.52 S, respectively, in good agreement with sedimentation coefficients predicted by hydrodynamic theory. The described approach can be applied to synthetic boundary and conventional loading experiments, and can be extended to analyze sedimentation data for both large and small macromolecules in order to define shape, heterogeneity, and state of association.

摘要

在溶剂和溶液平台不可用或为瞬态的条件下，利用沉降速度数据与Lamm方程的数值解直接拟合来获得单一溶质的沉降系数。计算演化以第一次实验扫描为初始条件，并采用非线性回归来获得沉降系数和扩散系数的最佳拟合值。研究了Lamm方程作为数据分析工具的一般特性。该方法应用于研究一组具有两亲性七肽重复序列、一般结构为Ac-YS-(AKEAAKE)nGAR-NH2（n = 2、3或4）的小肽。沉降速度分析表明存在单一沉降物种，其沉降系数（s(20,w)值）分别为0.37、0.45和0.52 S，与流体动力学理论预测的沉降系数高度一致。所描述的方法可应用于合成边界和传统加载实验，并且可以扩展到分析大小分子的沉降数据，以确定形状、异质性和缔合状态。