Saint Nathalie, Montserret Roland, Chipot Christophe, Penin François
Centre de Biochimie Structurale, Université de Montpellier I et II, Montpellier, France.
Methods Mol Biol. 2009;510:125-43. doi: 10.1007/978-1-59745-394-3_10.
The p7 membrane polypeptide from HCV is essential for virus infection. It exhibits ion-channel activity reported to be specifically blocked by various compounds. These properties make p7 an attractive candidate target for antiviral intervention to combat viral hepatitis C infection. In this context, in vitro functional analyses of isolated p7 coupled to structural characterization are critical for further understanding of the molecular mechanisms of p7 ion-channel activity and for the development of new antiviral drugs. We present here in vitro assays designed to purify synthetic p7 by RP-HPLC, to investigate its ion-channel properties by means of planar lipid-bilayer assays and patch-clamp recordings after reconstitution into liposomes, and to analyze its structural features by circular dichroism (CD), nuclear magnetic resonance (NMR), and molecular dynamics (MD).
丙型肝炎病毒(HCV)的p7膜多肽对病毒感染至关重要。它表现出离子通道活性,据报道各种化合物可特异性阻断该活性。这些特性使p7成为对抗丙型肝炎病毒感染的抗病毒干预的有吸引力的候选靶点。在此背景下,对分离的p7进行体外功能分析并结合结构表征,对于进一步了解p7离子通道活性的分子机制以及开发新的抗病毒药物至关重要。我们在此展示了旨在通过反相高效液相色谱(RP-HPLC)纯化合成p7、通过平面脂质双层分析和重组到脂质体后的膜片钳记录来研究其离子通道特性,以及通过圆二色性(CD)、核磁共振(NMR)和分子动力学(MD)分析其结构特征的体外实验。