Department of Chemistry, Institute for Biophysical Dynamics, University of Chicago, Chicago, IL 60637, USA.
Proc Natl Acad Sci U S A. 2011 Dec 27;108(52):20982-7. doi: 10.1073/pnas.1111202108. Epub 2011 Dec 8.
We have used chemical protein synthesis and advanced physical methods to probe dynamics-function correlations for the HIV-1 protease, an enzyme that has received considerable attention as a target for the treatment of AIDS. Chemical synthesis was used to prepare a series of unique analogues of the HIV-1 protease in which the flexibility of the "flap" structures (residues 37-61 in each monomer of the homodimeric protein molecule) was systematically varied. These analogue enzymes were further studied by X-ray crystallography, NMR relaxation, and pulse-EPR methods, in conjunction with molecular dynamics simulations. We show that conformational isomerization in the flaps is correlated with structural reorganization of residues in the active site, and that it is preorganization of the active site that is a rate-limiting factor in catalysis.
我们使用化学蛋白质合成和先进的物理方法来探究 HIV-1 蛋白酶的动态-功能相关性，该酶作为治疗艾滋病的靶点受到了广泛关注。化学合成被用于制备一系列独特的 HIV-1 蛋白酶类似物，其中“瓣”结构（每个同源二聚体蛋白分子单体的 37-61 位残基）的柔韧性被系统地改变。这些类似酶进一步通过 X 射线晶体学、NMR 弛豫和脉冲 EPR 方法以及分子动力学模拟进行研究。我们表明，瓣中的构象异构化与活性位点中残基的结构重排相关，并且活性位点的预组织是催化中的限速因素。
