Bonomi Massimiliano, Gervasio Francesco L, Tiana Guido, Provasi Davide, Broglia Ricardo A, Parrinello Michele
Computational Science, Department of Chemistry and Applied Biosciences, ETH Zürich, Lugano, Switzerland.
Biophys J. 2007 Oct 15;93(8):2813-21. doi: 10.1529/biophysj.107.106369. Epub 2007 Jun 15.
It has recently been shown that the highly protected segments 24-34 (S2) and 83-93 (S8) of each of the two 99-mers of human immunodeficiency virus type 1 protease play an essential role in the folding of the monomers, giving rise to the so-called (postcritical) folding nucleus ((FN) minimum condensation unit ensuring folding) when they dock. This scenario received further support from model calculations that demonstrated that the peptide p-S8, displaying an amino acid sequence identical to the corresponding (83-93) segment of the monomer, can be used to interfere with the formation of the FN and eventually to inhibit folding by docking the fragment 24-34. Experiments in vitro and in cells infected with ex vivo wild-type and multiresistant HIV isolates confirm that the inhibition power of p-S8 is robust. On the other hand, there is no direct evidence demonstrating the validity of the proposed mechanism of inhibition associated with p-S8. To shed light on this question and to provide the basis for the design of a molecule mimetic to p-S8, to be used as lead of an eventual drug against AIDS, we study, in this paper, with the help of all-atom simulations in explicit solvent and the novel method of metadynamics combined with parallel tempering: a), the free energy and the equilibrium structure of each of the peptides p-S2 and p-S8; b), the details of the docking mechanism of the two peptides and the free energy associated with this process. Whereas p-S8 is found to be well structured, p-S2 is rather flexible, wrapping itself around p-S8 to give rise to the FN, which is stabilized by three particular hydrogen bonds.
最近的研究表明,人类免疫缺陷病毒1型蛋白酶的两种99聚体中,每一种的高度保守片段24 - 34(S2)和83 - 93(S8)在单体折叠过程中起着至关重要的作用,当它们对接时会形成所谓的(临界后)折叠核((FN)确保折叠的最小凝聚单元)。这一设想得到了模型计算的进一步支持,该计算表明,肽p - S8(其氨基酸序列与单体相应的(83 - 93)片段相同)可用于干扰FN的形成,并最终通过对接片段24 - 34来抑制折叠。体外实验以及对感染了野生型和多抗性HIV分离株的细胞进行的实验证实,p - S8具有强大的抑制能力。另一方面,尚无直接证据证明与p - S8相关的抑制机制的有效性。为了阐明这个问题,并为设计一种模拟p - S8的分子提供基础,以便用作最终抗艾滋病药物的先导,我们在本文中借助显式溶剂中的全原子模拟以及结合平行回火的元动力学新方法,研究了:a)肽p - S2和p - S8各自的自由能和平衡结构;b)两种肽的对接机制细节以及与此过程相关的自由能。结果发现,p - S8结构良好,而p - S2相当灵活,它围绕p - S8缠绕形成FN,该FN通过三个特定的氢键得以稳定。