Pereira Martha Machado, Silva Bianca Alcântara, Pinto Marcia Ribeiro, Barreto-Bergter Eliana, dos Santos André Luis Souza
Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes (IMPPG), Centro de Ciências da Saúde (CCS), Bloco I, Universidade Federal do Rio de Janeiro (UFRJ), Ilha do Fundão, Rio de Janeiro, Brazil.
Mycopathologia. 2009 Jan;167(1):25-30. doi: 10.1007/s11046-008-9147-7. Epub 2008 Aug 23.
The conidia-mycelia transformation is an essential step during the life cycle of the fungal human pathogens of the Pseudallescheria boydii complex. In the present study, we have analyzed the protein and peptidase profiles in two distinct morphological stages, conidia and mycelia, of Scedosporium apiospermum sensu stricto. Proteins synthesized by the mycelia, migrating at the ranges of 62-48 and 22-18 kDa, were not detected from the conidial extract. Conidia produced a single cellular peptidase of 28 kDa able to digest copolymerized albumin, while mycelia yielded 6 distinct peptidases ranging from 90 to 28 kDa. All proteolytic enzymes were active at acidic pH and fully inhibited by 1,10-phenanthroline, characterizing these activities as metallo-type peptidases. Quantitative peptidase assay, using soluble albumin, showed a high metallopeptidase production in mycelial cells in comparison with conidia. The regulated expression of proteins and peptidases in different morphological stages of S. apiospermum represents a potential target for isolation of stage-specific markers for biochemical and immunological analysis.
分生孢子-菌丝体转变是博伊德假阿利什霉复合体真菌人类病原体生命周期中的一个重要步骤。在本研究中，我们分析了狭义腐皮镰刀菌两个不同形态阶段，即分生孢子和菌丝体中的蛋白质和肽酶谱。在分生孢子提取物中未检测到菌丝体合成的、迁移范围在62 - 48 kDa和22 - 18 kDa的蛋白质。分生孢子产生一种能够消化共聚白蛋白的28 kDa的单细胞肽酶，而菌丝体产生6种不同的肽酶，分子量范围从90 kDa到28 kDa。所有蛋白水解酶在酸性pH下具有活性，并被1,10 - 菲咯啉完全抑制，这些活性被鉴定为金属型肽酶。使用可溶性白蛋白进行的定量肽酶测定表明，与分生孢子相比，菌丝体细胞中金属肽酶的产量较高。狭义腐皮镰刀菌不同形态阶段蛋白质和肽酶的调控表达代表了分离用于生化和免疫分析的阶段特异性标志物的潜在靶点。
