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Optimizing protein complexes for crystal growth

机译:优化蛋白质复合物以促进晶体生长

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摘要

Many intracellular proteins do. not work on their own but rather in complex with small molecules, DNA, or other proteins. To gain a more fundamental understanding of protein interactions and their resulting functions, one requires a detailed structural model of relevant complexes. The first step in this challenge is to grow well-diffracting crystals. Three examples of protein complex crystallization will be discussed in detail below. In the first example, biophysical techniques such as fluorescence titration, isothermal titration calorimetry (ITC), and dynamic light scattering (DLS) are used to characterize the protein and assess the most suitable conditions for complex formation. The second example utilizes bioinformatic information and proteomic techniques to engineer constructs of the protein that are most favorable for crystallization. The final example uses NMR information for optimizing complex-forming conditions, which allowed the growth of better-diffracting complex crystals.
机译:许多细胞内蛋白都可以。不能单独工作,而是与小分子,DNA或其他蛋白质复合。为了对蛋白质相互作用及其产生的功能有更基本的了解,需要一种有关复合物的详细结构模型。这一挑战的第一步是生长衍射良好的晶体。蛋白质复合物结晶的三个例子将在下面详细讨论。在第一个示例中,使用生物物理技术(例如荧光滴定,等温滴定量热法(ITC)和动态光散射(DLS))表征蛋白质并评估最合适的复合物形成条件。第二个例子利用生物信息学信息和蛋白质组学技术来设计最适合结晶的蛋白质构建体。最后一个例子使用NMR信息优化了复合物的形成条件,从而使衍射更好的复合晶体得以生长。

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