Gupta Saurabh, Rao Atmakuri Ramakrishna, Varadwaj Pritish Kumar, De Sachinandan, Mohapatra Trilochan
Centre for Agricultural Bioinformatics, ICAR-Indian Agricultural Statistics Research Institute, Pusa, Library Avenue, New Delhi, 110012, India; Department of Bioinformatics, Indian Institute of Information Technology, Allahabad, 210012, India.
Centre for Agricultural Bioinformatics, ICAR-Indian Agricultural Statistics Research Institute, Pusa, Library Avenue, New Delhi, 110012, India.
PLoS One. 2015 Aug 27;10(8):e0136630. doi: 10.1371/journal.pone.0136630. eCollection 2015.
Heat shock protein 70 (HSP70) is an important chaperone, involved in protein folding, refolding, translocation and complex remodeling reactions under normal as well as stress conditions. However, expression of HSPA1A gene in heat and cold stress conditions associates with other chaperons and perform its function. Experimental structure for Camel HSP70 protein (cHSP70) has not been reported so far. Hence, we constructed 3D models of cHSP70 through multi- template comparative modeling with HSP110 protein of S. cerevisiae (open state) and with HSP70 protein of E. coli 70kDa DnaK (close state) and relaxed them for 100 nanoseconds (ns) using all-atom Molecular Dynamics (MD) Simulation. Two stable conformations of cHSP70 with Substrate Binding Domain (SBD) in open and close states were obtained. The collective mode analysis of different transitions of open state to close state and vice versa was examined via Principal Component Analysis (PCA) and Minimum Distance Matrix (MDM). The results provide mechanistic representation of the communication between Nucleotide Binding Domain (NBD) and SBD to identify the role of sub domains in conformational change mechanism, which leads the chaperone cycle of cHSP70. Further, residues present in the chaperon functioning site were also identified through protein-peptide docking. This study provides an overall insight into the inter domain communication mechanism and identification of the chaperon binding cavity, which explains the underlying mechanism involved during heat and cold stress conditions in camel.
热休克蛋白70(HSP70)是一种重要的伴侣蛋白,在正常以及应激条件下参与蛋白质折叠、重折叠、转运和复合物重塑反应。然而,HSPA1A基因在热应激和冷应激条件下的表达与其他伴侣蛋白相关联并发挥其功能。迄今为止,骆驼HSP70蛋白(cHSP70)的实验结构尚未见报道。因此,我们通过与酿酒酵母的HSP110蛋白(开放状态)和大肠杆菌70kDa DnaK的HSP70蛋白(关闭状态)进行多模板比较建模构建了cHSP70的三维模型,并使用全原子分子动力学(MD)模拟将其松弛100纳秒(ns)。获得了cHSP70在开放和关闭状态下具有底物结合结构域(SBD)的两种稳定构象。通过主成分分析(PCA)和最小距离矩阵(MDM)研究了开放状态到关闭状态以及反之的不同转变的集体模式分析。结果提供了核苷酸结合结构域(NBD)和SBD之间通讯的机制表示,以确定亚结构域在构象变化机制中的作用,这导致了cHSP70的伴侣循环。此外,还通过蛋白质 - 肽对接鉴定了伴侣蛋白功能位点中存在的残基。这项研究提供了对结构域间通讯机制的全面洞察以及伴侣蛋白结合腔的鉴定,这解释了骆驼在热应激和冷应激条件下所涉及的潜在机制。