As the number of high-resolution structures of membrane proteins continues to rise, so has the necessity for techniques to link this structural information to protein function. In the case of transporters, function is achieved via coupling of conformational changes to substrate binding and release. Static structural data alone cannot convey information on these protein movements, but it can provide a high-resolution foundation on which to interpret lower resolution data obtained by complementary approaches. Here, we review selected biochemical and spectroscopic methods for assessing transporter conformational change. In addition to more traditional techniques, we present ¹⁹F-NMR as an attractive method for characterizing conformational change in transporters of known structure. Using biosynthetic labeling, multiple, non-perturbing fluorine-labeled amino acids can be incorporated throughout a protein to serve as reporters of conformational change. Such flexibility in labeling allows characterization of movement in protein regions that may not be accessible via other methods.

中文翻译：

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在晶体之外思考：检查转运蛋白构象变化的补充方法。

随着膜蛋白高分辨率结构的数量不断增加，将这种结构信息与蛋白功能联系起来的技术的必要性也越来越高。对于转运蛋白，通过构象变化与底物结合和释放的偶联来实现功能。静态结构数据本身无法传达有关这些蛋白质运动的信息，但是它可以提供高分辨率的基础，在该基础上可以解释通过互补方法获得的较低分辨率的数据。在这里，我们审查了选定的生化和光谱方法，以评估转运蛋白的构象变化。除更传统的技术外，我们提出的1 F-NMR是表征已知结构转运蛋白构象变化的一种有吸引力的方法。使用生物合成标记，可以在整个蛋白质中掺入非干扰性的氟标记氨基酸，以作为构象变化的报告子。标记的这种灵活性允许表征蛋白质区域中的运动，而这些区域可能无法通过其他方法获得。