Abstract：
In the realm of parasitology, autophagy has emerged as a critical focal point, particularly in combating Leishmaniasis. Central to this endeavour is the recognition of the protein ATG8 as pivotal for the survival and infectivity of the parasitic organism Leishmania major, thereby making it a potential target for therapeutic intervention. Consequently, there is a pressing need to delve into the structural characteristics of ATG8 to facilitate the design of effective drugs. In this study, our efforts centered on the purification of ATG8 from Leishmania major, which enabled novel insights into its structural features through meticulous spectroscopic analysis. We aimed to comprehensively assess the stability and behaviour of ATG8 in the presence of various denaturants, including urea, guanidinium chloride, and SDS-based chemicals. Methodically, our approach included secondary structural analysis utilizing CD spectroscopy, which not only validated but also augmented computationally predicted structures of ATG8 reported in previous investigations. Remarkably, our findings unveiled that the purified ATG8 protein retained its folded conformation, exhibiting the anticipated secondary structure. Moreover, our exploration extended to the influence of lipids on ATG8 stability, yielding intriguing revelations. We uncovered a nuanced perspective suggesting that targeting both the lipid composition of Leishmania major and ATG8 could offer a promising strategy for future therapeutic approaches in combating leishmaniasis. Collectively, our study underscores the importance of understanding the structural intricacies of ATG8 in driving advancements towards the development of targeted therapies against Leishmaniasis, thereby providing a foundation for future investigations in this field.
摘要：
在寄生虫学领域，自噬已经成为一个关键的焦点，特别是在对抗利什曼病方面。这项努力的核心是认识到ATG8蛋白对于主要寄生生物利什曼原虫的生存和传染性至关重要，从而使其成为治疗干预的潜在目标。因此,迫切需要深入研究ATG8的结构特征，以促进有效药物的设计。在这项研究中,我们的努力集中在从利什曼原虫中纯化ATG8，这使得通过细致的光谱分析对其结构特征有了新的见解。我们旨在全面评估ATG8在各种变性剂存在下的稳定性和行为，包括尿素,氯化胍,和基于SDS的化学品。有条不紊地，我们的方法包括利用CD光谱学的二级结构分析，这不仅验证了，而且增强了先前研究中报道的ATG8的计算预测结构。值得注意的是,我们的发现揭示了纯化的ATG8蛋白保留了其折叠构象，表现出预期的二级结构。此外，我们的探索扩展到脂质对ATG8稳定性的影响，产生有趣的启示。我们发现了一个微妙的观点，表明靶向利什曼原虫和ATG8的脂质组成可以为未来对抗利什曼病的治疗方法提供有希望的策略。总的来说，我们的研究强调了了解ATG8的结构复杂性在推动发展针对利什曼病的靶向治疗方面的重要性，从而为该领域的未来调查奠定了基础。
