Rapid Distinction of Intracellular and Extracellular Proteins Using NMR Diffusion Measurements

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• 作者：Christopher A. Waudby ; Mick D. Mantle ; Lisa D. Cabrita ; Lynn F. Gladden ; Christopher M. Dobson ; John Christodoulou
• 刊名：The Journal of the American Chemical Society
• 出版年：2012
• 出版时间：July 18, 2012
• 年：2012
• 卷：134
• 期：28
• 页码：11312-11315
• 全文大小：270K
• 年卷期：v.134,no.28(July 18, 2012)
• ISSN：1520-5126

文摘

In-cell NMR spectroscopy offers a unique opportunity to begin to investigate the structures, dynamics, and interactions of molecules within their functional environments. An essential aspect of this technique is to define whether observed signals are attributable to intracellular species rather than to components of the extracellular medium. We report here the results of NMR measurements of the diffusion behavior of proteins expressed within bacterial cells, and find that these experiments provide a rapid and nondestructive probe of localization within cells and can be used to determine the size of the confining compartment. We show that diffusion can also be exploited as an editing method to eliminate extracellular species from high-resolution multidimensional spectra, and should be applicable to a wide range of problems. This approach is demonstrated here for a number of protein systems, using both ¹⁵N and ¹³C (methyl-TROSY) based acquisition.