Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal de Goiás, 74001-970, Goiânia, Goiás, Brazil.
Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, 85819-110, Cascavel, Paraná, Brazil.
Microbes Infect. 2019 Oct-Nov;21(8-9):401-411. doi: 10.1016/j.micinf.2019.03.002. Epub 2019 Apr 2.
Paracoccidioidomycosis (PCM), caused by thermodimorphic fungi of the Paracoccidioides genus, is a systemic disorder that involves the lungs and other organs. The adherence of pathogenic microorganisms to host tissues is an essential event in the onset of colonization and spread. The host-pathogen interaction is a complex interplay between the defense mechanisms of the host and the efforts of pathogenic microorganisms to colonize it. Therefore, the identification of fungi proteins interacting with host proteins is an important step understanding the survival strategies of the fungus within the host. In this paper, we used affinity chromatography based on surface proteomics (ACSP) to investigate the interactions of pathogen proteins with host surface molecules. Paracoccidioides lutzii extracts enriched of surface proteins were captured by chromatographic resin, which was immobilized with macrophage cell surface proteins, and identified by mass spectrometry. A total of 215 proteins of P. lutzii were identified interacting with macrophage proteins. In silico analysis classified those proteins according to the presence of sites for N- and O-glycosylation and secretion by classical and non-classical pathways. Serine proteinase (SP) and fructose-1,6-bisphosphate aldolase (FBA) were identified in our proteomics analysis. Immunolocalization assay and flow cytometry both showed an increase in the expression of these two proteins during host-pathogen interaction.
球孢子菌病（PCM），由球腔菌属的热双相真菌引起，是一种涉及肺部和其他器官的系统性疾病。致病微生物对宿主组织的黏附是定植和传播开始的一个重要事件。宿主-病原体的相互作用是宿主防御机制与致病微生物定植它的努力之间复杂的相互作用。因此，鉴定与宿主蛋白相互作用的真菌蛋白是了解真菌在宿主内生存策略的重要步骤。在本文中，我们使用基于表面蛋白质组学的亲和层析（ACSP）来研究病原体蛋白与宿主表面分子的相互作用。用固定有巨噬细胞表面蛋白的色谱树脂捕获富含表面蛋白的副球孢子菌提取物，并通过质谱进行鉴定。鉴定出 215 种与巨噬细胞蛋白相互作用的副球孢子菌蛋白。通过计算机分析，根据 N 和 O 糖基化位点的存在以及经典和非经典途径的分泌，对这些蛋白进行了分类。丝氨酸蛋白酶（SP）和果糖-1,6-二磷酸醛缩酶（FBA）在我们的蛋白质组学分析中被鉴定出来。免疫定位分析和流式细胞术都显示了这两种蛋白在宿主-病原体相互作用过程中表达增加。
