Yamazaki T, Nicholson L K, Torchia D A, Stahl S J, Kaufman J D, Wingfield P T, Domaille P J, Campbell-Burk S
Bone Research Branch, National Institute of Dental Research, NIH, Bethesda, MD 20892.
Eur J Biochem. 1994 Jan 15;219(1-2):707-12. doi: 10.1111/j.1432-1033.1994.tb19987.x.
We report comprehensive NMR studies in solution of the human-immunodeficiency-virus (HIV)-1 protease. Stable solutions of the protease were obtained by complexing the protein to a designed cyclic urea inhibitor DMP 323. A variety of triple-resonance experiments provided essentially complete 1H, 13C and 15N NMR signal assignments of the protease. These assignments, together with short-range NOE constraints, coupling constants and hydrogen-exchange data, yielded the secondary structure of the protease in solution. The results reported herein open the way to the determination of the high-resolution three-dimensional solution structures of protease/inhibitor complexes, as well as to studies of protease dynamics and solvent interactions.
我们报告了对人类免疫缺陷病毒（HIV）-1蛋白酶在溶液中的全面核磁共振（NMR）研究。通过将该蛋白质与设计的环脲抑制剂DMP 323复合，获得了蛋白酶的稳定溶液。各种三共振实验基本上完成了蛋白酶的1H、13C和15N NMR信号归属。这些归属，连同短程核Overhauser效应（NOE）约束、耦合常数和氢交换数据，得出了溶液中蛋白酶的二级结构。本文报道的结果为确定蛋白酶/抑制剂复合物的高分辨率三维溶液结构以及研究蛋白酶动力学和溶剂相互作用开辟了道路。
