Jiang Wenqi, Wu Lian, Shen Xiangchun, Fu Qingshan Bill
State Key Laboratory of Discovery and Utilization of Functional Components in Traditional Chinese Medicine, School of Pharmaceutical Sciences, Guizhou Medical University, Guizhou, 561113, China.
Shanghai Institute of Materia Medica, Zhongshan Institute for Drug Discovery, Chinese Academy of Sciences, Zhongshan, 528400, China.
Protein Pept Lett. 2025;32(5):376-386. doi: 10.2174/0109298665390494250513110604.
Human papillomavirus type 16 (HPV16) is implicated in various malignancies. The virus enters host cells through endocytosis, during which the minor capsid protein L2 interacts with the S100A10 subunit of the annexin A2 heterotetramer (A2t) on the host cell membrane. This interaction is critical for facilitating HPV entry and subsequent infection of human cells. Therefore, examining the interaction between the L2 protein and S100A10 is crucial for advancing our understanding of the mechanisms by which HPV16 infiltrates cells.
The cell-free expression (CFE) system was investigated for L2 purification. The structure of L2 was characterized and its interaction with S100A10 was explored.
The L2 protein was expressed using a CFE expression system, and its expression was verified via Western blotting. L2 was further purified through size-exclusion chromatography (SEC), and its structural features were preliminarily assessed using transmission electron microscopy (TEM) and circular dichroism (CD) spectroscopy. Additionally, surface plasmon resonance (SPR) was employed to analyze the interaction between L2 and S100A10.
Western blotting confirmed the successful expression of L2. TEM and CD provided preliminary structural observations of L2, and SPR measurements yielded precise kinetic parameters for the interaction between L2 and S100A10.
In this study, we successfully expressed the HPV16 L2 protein using a cell-free protein expression system. Preliminary structural analysis using TEM and CD revealed key structural features of L2. Furthermore, SPR analysis provided detailed kinetic parameters for its interaction with S100A10. These findings provide more details on understanding L2's structural features, with broader implications for antipathogen studies.
16型人乳头瘤病毒（HPV16）与多种恶性肿瘤有关。该病毒通过内吞作用进入宿主细胞，在此过程中，次要衣壳蛋白L2与宿主细胞膜上膜联蛋白A2异源四聚体（A2t）的S100A10亚基相互作用。这种相互作用对于促进HPV进入及随后感染人类细胞至关重要。因此，研究L2蛋白与S100A10之间的相互作用对于深入了解HPV16侵入细胞的机制至关重要。
研究无细胞表达（CFE）系统用于L2纯化的情况。对L2的结构进行表征，并探索其与S100A10的相互作用。
使用CFE表达系统表达L2蛋白，并通过蛋白质免疫印迹法验证其表达。通过尺寸排阻色谱法（SEC）进一步纯化L2，并使用透射电子显微镜（TEM）和圆二色性（CD）光谱对其结构特征进行初步评估。此外，采用表面等离子体共振（SPR）分析L2与S100A10之间的相互作用。
蛋白质免疫印迹法证实L2成功表达。TEM和CD提供了L2的初步结构观察结果，SPR测量得出了L2与S100A10相互作用的精确动力学参数。
在本研究中，我们使用无细胞蛋白质表达系统成功表达了HPV16 L2蛋白。使用TEM和CD进行的初步结构分析揭示了L2的关键结构特征。此外，SPR分析提供了其与S100A10相互作用的详细动力学参数。这些发现为理解L2的结构特征提供了更多细节，在抗病原体研究中具有更广泛的意义。
