Cooley J W, Abdine A, Brown M, Chavez J, Lada B, JiJi R D, Ubarretxena-Belandia I
University of Missouri, Columbia, MO, United States.
Icahn School of Medicine at Mount Sinai, New York, NY, United States.
Methods Enzymol. 2017;584:207-228. doi: 10.1016/bs.mie.2016.10.030. Epub 2016 Dec 9.
We present a new method based on deep-UV resonance Raman spectroscopy to determine the backbone conformation of intramembrane protease substrates. The classical amide vibrational modes reporting on the conformation of just the transmembrane region of the substrate can be resolved from solvent exchangeable regions outside the detergent micelle by partial deuteration of the solvent. In the presence of isotopically triple-labeled intramembrane protease, these amide modes can be accurately measured to monitor the transmembrane conformation of the substrate during intramembrane proteolysis.
我们提出了一种基于深紫外共振拉曼光谱的新方法，用于确定膜内蛋白酶底物的主链构象。通过对溶剂进行部分氘代，可以将仅报告底物跨膜区域构象的经典酰胺振动模式与去污剂胶束外的溶剂可交换区域区分开来。在存在同位素三重标记的膜内蛋白酶的情况下，可以准确测量这些酰胺模式，以监测膜内蛋白水解过程中底物的跨膜构象。
