Kessenbrock Mareike, Groth Georg
Institute of Biochemical Plant Physiology, Heinrich-Heine University Düsseldorf, Universitätsstraße 1, D-40225, Düsseldorf, Germany.
Methods Mol Biol. 2017;1573:141-159. doi: 10.1007/978-1-4939-6854-1_12.
Circular dichroism (CD) spectroscopy is an invaluable technique to analyze secondary structure and functional folding of recombinant purified proteins. CD spectroscopy can also be applied to detect changes in protein secondary structure related to the pH or redox conditions found in different cellular compartments or to the interaction with other molecules. Another biophysical technique to monitor conformational changes and interaction with small molecule ligands or biological macromolecules is protein fluorescence spectroscopy making use of the aromatic amino acid tryptophan as a sensitive intrinsic fluorescent probe. Here, we describe the application of CD and tryptophan fluorescence spectroscopy to study soluble and membrane proteins of the ethylene signaling pathway.
圆二色性（CD）光谱是分析重组纯化蛋白质二级结构和功能折叠的一项宝贵技术。CD光谱还可用于检测与不同细胞区室中发现的pH或氧化还原条件相关的蛋白质二级结构变化，或与其他分子相互作用引起的变化。另一种监测构象变化以及与小分子配体或生物大分子相互作用的生物物理技术是蛋白质荧光光谱，它利用芳香族氨基酸色氨酸作为灵敏的内在荧光探针。在此，我们描述了CD和色氨酸荧光光谱在研究乙烯信号通路的可溶性和膜蛋白中的应用。
