Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, ICB II, Campus II, Universidade Federal de Goiás, 74001-970 Goiânia, Goiás, Brazil.
Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, ICB II, Campus II, Universidade Federal de Goiás, 74001-970 Goiânia, Goiás, Brazil; Laboratório Interdisciplinar de Biologia, Universidade Estadual de Goiás, Itapuranga, Goiás, Brazil.
Biochim Biophys Acta Proteins Proteom. 2017 Nov;1865(11 Pt A):1304-1314. doi: 10.1016/j.bbapap.2017.08.016. Epub 2017 Aug 24.
Paracoccidioidomycosis is an important systemic mycosis caused by thermodimorphic fungi of the Paracoccidioides genus. During the infective process, the cell wall acts at the interface between the fungus and the host. In this way, the cell wall has a key role in growth, environment sensing and interaction, as well as morphogenesis of the fungus. Since the cell wall is absent in mammals, it may present molecules that are described as target sites for new antifungal drugs. Despite its importance, up to now few studies have been conducted employing proteomics in for the identification of cell wall proteins in Paracoccidioides spp. Here, a detailed proteomic approach, including cell wall-fractionation coupled to NanoUPLC-MS, was used to study and compare the cell wall fractions from Paracoccidioides lutzii mycelia and yeast cells. The analyzed samples consisted of cell wall proteins extracted by hot SDS followed by extraction by mild alkali. In summary, 512 proteins constituting different cell wall fractions were identified, including 7 predicted GPI-dependent cell wall proteins that are potentially involved in cell wall metabolism. Adhesins previously described in Paracoccidioides spp. such as enolase, glyceraldehyde-3-phosphate dehydrogenase were identified. Comparing the proteins in mycelium and yeast cells, we detected some that are common to both fungal phases, such as Ecm33, and some specific proteins, as glucanase Crf1. All of those proteins were described in the metabolism of cell wall. Our study provides an important elucidation of cell wall composition of fractions in Paracoccidioides, opening a way to understand the fungus cell wall architecture.
球孢子菌病是一种由球腔菌属的嗜热丝状真菌引起的重要系统性真菌病。在感染过程中，细胞壁位于真菌和宿主之间的界面上。因此，细胞壁在真菌的生长、环境感应和相互作用以及形态发生中起着关键作用。由于哺乳动物中不存在细胞壁，因此它可能具有被描述为新型抗真菌药物靶标的分子。尽管细胞壁很重要，但迄今为止，利用蛋白质组学研究球腔菌属细胞壁蛋白的研究很少。在这里，采用详细的蛋白质组学方法，包括细胞壁分离与纳升超高效液相色谱-串联质谱（NanoUPLC-MS）联用，研究并比较了球腔菌属丝状菌和酵母细胞的细胞壁部分。分析的样品由热 SDS 提取的细胞壁蛋白组成，然后用温和的碱提取。总之，鉴定了构成不同细胞壁部分的 512 种蛋白质，包括 7 种预测的 GPI 依赖性细胞壁蛋白，它们可能参与细胞壁代谢。在球腔菌属中先前描述的粘附素，如烯醇酶、甘油醛-3-磷酸脱氢酶也被鉴定出来。比较丝状菌和酵母细胞中的蛋白质，我们检测到一些在两个真菌相中都存在的共同蛋白，如 Ecm33，还有一些特定的蛋白，如葡聚糖酶 Crf1。所有这些蛋白质都参与了细胞壁的代谢。我们的研究为球腔菌属各部分细胞壁的组成提供了重要的阐明，为理解真菌细胞壁结构开辟了途径。
