Freitas E Silva Kleber Santiago, Melo Lima Raisa, de Sousa Lima Patrícia, Cristiane Baeza Lilian, Alves da Silva Roosevelt, Maria de Almeida Soares Célia, Pereira Maristela
Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal de Goiás, Goiânia 74690-900, Goiás, Brazil.
Laboratório de Microbiologia Experimental, Centro de Ciências Médicas e Farmacêuticas, Universidade Estadual do Oeste do Paraná, Cascavel 85819-170, Paraná, Brazil.
J Fungi (Basel). 2020 Nov 23;6(4):309. doi: 10.3390/jof6040309.
Systemic mycosis is a cause of death of immunocompromised subjects. The treatment directed to evade fungal pathogens shows severe limitations, such as time of drug exposure and side effects. The paracoccidioidomycosis (PCM) treatment depends on the severity of the infection and may last from months to years.
To analyze the main interactions of isocitrate lyase (ICL) regarding the energetic metabolism through affinity chromatography, we performed blue native PAGE and co-immunoprecipitation to identify ICL interactions. We also performed in silico analysis by homology, docking, hot-spot prediction and contact preference analysis to identify the conformation of ICL complexes.
ICL interacted with 18 proteins in mycelium, 19 in mycelium-to-yeast transition, and 70 in yeast cells. Thirty complexes were predicted through docking and contact preference analysis. ICL has seven main regions of interaction with protein partners.
ICL seems to interfere with energetic metabolism of , regulating aerobic and anaerobic metabolism as it interacts with proteins from glycolysis, gluconeogenesis, TCA and methylcitrate cycles, mainly through seven hot-spot residues.
系统性真菌病是免疫功能低下患者的死亡原因之一。针对逃避真菌病原体的治疗存在严重局限性，如药物暴露时间和副作用。副球孢子菌病（PCM）的治疗取决于感染的严重程度，可能持续数月至数年。
为了通过亲和色谱分析异柠檬酸裂解酶（ICL）在能量代谢方面的主要相互作用，我们进行了蓝色非变性聚丙烯酰胺凝胶电泳（blue native PAGE）和免疫共沉淀以鉴定ICL的相互作用。我们还通过同源性、对接、热点预测和接触偏好分析进行了计算机模拟分析，以鉴定ICL复合物的构象。
ICL在菌丝体中与18种蛋白质相互作用，在菌丝体向酵母转变过程中与19种蛋白质相互作用，在酵母细胞中与70种蛋白质相互作用。通过对接和接触偏好分析预测了30种复合物。ICL与蛋白质伴侣有七个主要相互作用区域。
ICL似乎干扰了[此处原文可能有误，推测应为某种真菌的名称]的能量代谢，通过主要与糖酵解、糖异生、三羧酸循环和甲基柠檬酸循环中的蛋白质相互作用，主要通过七个热点残基来调节有氧和无氧代谢。
