Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran.
Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran.
Int J Biol Macromol. 2017 Dec;105(Pt 1):66-73. doi: 10.1016/j.ijbiomac.2017.06.113. Epub 2017 Jul 1.
Renilla luciferase (Rluc) from Renilla reniformis is an appropriate protein reporter for the detection of specific molecular targets due to its bioluminescent feature, although its relatively low stability limits the application. To investigate the effects of trehalose and sucrose as chemical chaperones on the kinetic stability of Rluc, we assayed the activity of the enzyme in the presence of these additives at high temperatures and to comprehend the mechanism of stability, molecular dynamic (MD) simulation was carried out. In the presence of trehalose a thermostabilizing effect which was considerable in comparison with other systems was observed. It is proposed that a wide radial like network of trehalose molecules supports α-helix structures that are located in the N-terminus and C-terminus of the protein. However, in the water simulation box, these helices alter to instable structures at high temperatures. Reduction of the fluctuation of these helices in the presence of trehalose molecules, may prevent the protein from unfolding and increase its shelf life.
海肾荧光素酶（Rluc）来自海肾，由于其生物发光特性，是检测特定分子靶标的合适蛋白报告基因，但其相对较低的稳定性限制了其应用。为了研究海藻糖和蔗糖作为化学伴侣对 Rluc 动力学稳定性的影响，我们在高温下检测了在这些添加剂存在下酶的活性，并为了理解稳定性的机制，进行了分子动力学（MD）模拟。在海藻糖存在下，观察到与其他系统相比相当显著的热稳定效应。据推测，海藻糖分子的宽辐射状网络支持位于蛋白质的 N 端和 C 端的α-螺旋结构。然而，在水模拟盒中，这些螺旋在高温下会变成不稳定的结构。海藻糖分子的存在减少了这些螺旋的波动，可能防止了蛋白质的展开并增加了其保质期。
