Curley R W, Panigot M J, Hansen A P, Fesik S W
Ohio State University, Columbus 43210.
J Biomol NMR. 1994 May;4(3):335-40. doi: 10.1007/BF00179344.
A method is described for stereospecifically assigning the alpha-protons of glycine residues in proteins. The approach involves the stereospecific deuteration and 15N labeling of glycine and subsequent selective incorporation of this residue into the protein. The stereospecific assignments of the glycine alpha-protons are obtained from a comparison of a 3D 15N-resolved TOCSY spectrum of the uniformly 15N-labeled protein with a 2D/3D 15N-edited TOCSY spectrum of the protein, containing the stereospecifically deuterated and 15N-labeled glycine. The approach is demonstrated by stereospecifically assigning the glycine alpha-protons of the FK506 binding protein when bound to the immunosuppressant ascomycin.
本文描述了一种对蛋白质中甘氨酸残基的α-质子进行立体专一性归属的方法。该方法包括对甘氨酸进行立体专一性氘代和¹⁵N标记，随后将该残基选择性掺入蛋白质中。通过比较均匀¹⁵N标记蛋白质的3D¹⁵N分辨TOCSY谱与含有立体专一性氘代和¹⁵N标记甘氨酸的蛋白质的2D/3D¹⁵N编辑TOCSY谱，获得甘氨酸α-质子的立体专一性归属。当FK506结合蛋白与免疫抑制剂子囊霉素结合时，通过对其甘氨酸α-质子进行立体专一性归属证明了该方法。
