Wenqi Jiang, Lian Wu, Xiangchun Shen, Qingshan Bill Fu
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引用次数: 0
Abstract
Background: 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.
Objective: The cell-free expression (CFE) system was investigated for L2 purification. The structure of L2 was characterized and its interaction with S100A10 was explored.
Methods: 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.
Results: 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.
Conclusion: 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.
期刊介绍：
Protein & Peptide Letters publishes letters, original research papers, mini-reviews and guest edited issues in all important aspects of protein and peptide research, including structural studies, advances in recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, and drug design. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallization and preliminary structure determination of biologically important proteins are considered only if they include significant new approaches or deal with proteins of immediate importance, and preliminary structure determinations of biologically important proteins. Purely theoretical/review papers should provide new insight into the principles of protein/peptide structure and function. Manuscripts describing computational work should include some experimental data to provide confirmation of the results of calculations.
Protein & Peptide Letters focuses on:
Structure Studies
Advances in Recombinant Expression
Drug Design
Chemical Synthesis
Function
Pharmacology
Enzymology
Conformational Analysis
Immunology
Biotechnology
Protein Engineering
Protein Folding
Sequencing
Molecular Recognition
Purification and Analysis
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