ANALYTICAL ULTRACENTRIFUGATION OF COMPLEX MACROMOLECULAR SYSTEMS
ANALYTICAL ULTRACENTRIFUGATION OF COMPLEX MACROMOLECULAR SYSTEMS
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DOI：
10.1021/bi00249a001
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发表时间：
1994-11-15
期刊：
影响因子：
2.9
通讯作者：
DEMELER, B
中科院分区：
文献类型：
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作者：
HANSEN, JC;LEBOWITZ, J;DEMELER, B
分析型超重力仪能直接测量大分子的沉降系数s、扩散系数D和分子量M，这些大分子的大小从几千道尔顿到几千万道尔顿不等。当与适当的数据分析方法相结合时，它可用于检查样品纯度，检测和表征构象变化，确定亚基化学计量，表征大分子复合物的组装和拆卸机制，以及测量缔合系统的平衡常数和热力学参数。因此，分析超离心是一个强大的工具，应该包括在剧目的任何研究人员感兴趣的生物大分子的结构-功能关系。尽管如此，在过去的20年里，分析性超集中化的使用急剧下降[参见Schachman（1992）的历史回顾]。这几乎完全可以追溯到在此期间无法使用功能性仪器的事实。然而，最近一种新的最先进的分析超浓缩仪Beckman XL-A的可用性扭转了这一趋势。重要的是，在大多数情况下，XL-A的能力大大超过了以前版本的分析型超离心分析仪Beckman Model E。因此，分析性的超离心不是简单地重生了，而是经历了真正的蜕变。该观点描述了这一变化的那些方面，这些方面最深刻地影响了NIH赠款GM 45916和RR 08352（JCH）和P30 AI 27767（JL）支持的许多项目。
An analytical ultracentrifuge is capable of directly measur-ing the sedimentationcoefficient, s, the diffusion coefficient, D, and the molecular mass, M, of macromolecules ranging in size from several thousand to tens of millions of daltons. When coupled with appropriate data analysis methodologies, it can be used to examine sample purity, detect and characterize conformational changes, determine subunit stoichiometries, characterize assembly and disassembly mechanisms of macromolecular complexes, and measure equilibrium constants and thermodynamic parameters for associating systems. As such, analytical ultracentrifugation is a powerful tool that should be included in the repertoire of any researcher interested in the structure—function relationships of biological macromolecules. Nonetheless, there has been a drastic decline in the use of analytical ultracentrifugation over the last 20 years [see Schachman (1992) for a historical review]. This can be traced almost solely to the fact that functional instrumentation has been unavailable during this period. However, the recent avail-ability of a new state-of-the-art analytical ultracentrifuge, the Beckman XL-A, has reversed this trend. Importantly, the capabilities of the XL-A in most cases have greatly exceeded those of the previous version of the analytical ultracentrifuge, the Beckman Model E. Consequently, analytical ultracen-trifugation has not simply been reborn, but has undergone a true metamorphosis. This perspective describes those aspects of this change that most profoundly impact the many t Supported by NIH Grants GM45916 and RR08352 (JCH) and P30AI27767 (JL).