Marcink Tara C, Koppisetti Rama K, Fulcher Yan G, Van Doren Steven R
Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, MO, 65211, USA.
Department of Medical Microbiology and Immunology, Life Sciences Center, University of Missouri, Columbia, MO, 65211, USA.
Methods Mol Biol. 2017;1579:61-86. doi: 10.1007/978-1-4939-6863-3_5.
Peripheral binding of proteins to lipid bilayers is critical not only in intracellular signaling but also in metalloproteinase shedding of signaling proteins from cell surfaces. Assessment of how proteins recognize fluid bilayers peripherally using crystallography or structure-based predictions has been important but incomplete. Assay of dynamic protein-bilayer interactions in solution has become feasible and reliable using paramagnetic NMR and site-directed fluor labeling. Details of preparations and assay protocols for these spectroscopic measurements of bilayer proximity or contact, respectively, are described.
蛋白质与脂质双层的外周结合不仅在细胞内信号传导中至关重要,而且在信号蛋白从细胞表面的金属蛋白酶裂解过程中也起着关键作用。利用晶体学或基于结构的预测来评估蛋白质如何在外周识别流体双层一直很重要,但并不完整。使用顺磁核磁共振和定点荧光标记,在溶液中测定动态蛋白质-双层相互作用已变得可行且可靠。本文分别描述了用于这些双层接近度或接触的光谱测量的制备细节和测定方案。