Department of Molecular and Cellular Biology, University of Guelph, Guelph N1G 2W1, Ontario, Canada.
Bruker Ltd., Milton L9T 6P4, Ontario, Canada.
J Proteome Res. 2024 Sep 6;23(9):3917-3932. doi: 10.1021/acs.jproteome.4c00255. Epub 2024 Aug 14.
Accurate and reliable detection of fungal pathogens presents an important hurdle to manage infections, especially considering that fungal pathogens, including the globally important human pathogen, , have adapted diverse mechanisms to survive the hostile host environment and moderate virulence determinant production during coinfections. These pathogen adaptations present an opportunity for improvements (e.g., technological and computational) to better understand the interplay between a host and a pathogen during disease to uncover new strategies to overcome infection. In this study, we performed comparative proteomic profiling of an in vitro coinfection model across a range of fungal and bacterial burden loads in macrophages. Comparing data-dependent acquisition and data-independent acquisition enabled with parallel accumulation serial fragmentation technology, we quantified changes in dual-perspective proteome remodeling. We report enhanced and novel detection of pathogen proteins with data-independent acquisition-parallel accumulation serial fragmentation (DIA-PASEF), especially for fungal proteins during single and dual infection of macrophages. Further characterization of a fungal protein detected only with DIA-PASEF uncovered a novel determinant of fungal virulence, including altered capsule and melanin production, thermotolerance, and macrophage infectivity, supporting proteomics advances for the discovery of a novel putative druggable target to suppress pathogenicity.
准确可靠地检测真菌病原体是管理感染的一个重要障碍，特别是考虑到真菌病原体，包括全球重要的人类病原体 ，已经适应了多种机制来在宿主环境中生存和在合并感染时适度产生毒力决定因子。这些病原体的适应为改进（例如，技术和计算）提供了机会，以更好地了解宿主和病原体在疾病过程中的相互作用，从而发现克服感染的新策略。在这项研究中，我们在体外共感染模型中进行了比较蛋白质组学分析，研究了巨噬细胞中真菌和细菌负担的一系列变化。通过比较基于数据的采集和基于平行累积串联碎裂技术的独立数据采集，我们定量了双视角蛋白质组重塑的变化。我们报告了独立数据采集-平行累积串联碎裂（DIA-PASEF）对病原体蛋白的增强和新颖检测，特别是在巨噬细胞单感染和双感染期间对真菌蛋白的检测。对仅用 DIA-PASEF 检测到的一种真菌蛋白的进一步表征揭示了一种新的真菌毒力决定因子，包括改变的荚膜和黑色素产生、耐热性和巨噬细胞感染性，支持蛋白质组学在发现新型潜在可药物靶标以抑制致病性方面的进展。
