Satala Dorota, Satala Grzegorz, Kulig Kamila, Karkowska-Kuleta Justyna, Kozik Andrzej, Rapala-Kozik Maria
Department of Comparative Biochemistry and Bioanalytics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, Kraków, 30-387, Poland.
Department of Medicinal Chemistry, Maj Institute of Pharmacology, Polish Academy of Sciences, Smetna 12, Kraków, Poland.
Sci Rep. 2025 Sep 1;15(1):32115. doi: 10.1038/s41598-025-15577-6.
Candida glabrata (currently classified as Nakaseomyces glabratus) is an opportunistic yeast-like fungus that causes infections in humans, with limited treatment options due to resistance to antifungal drugs. In contrast to C. albicans, which produces secreted aspartic proteases (Saps) involved in pathogenicity, C. glabrata expresses a distinct group of cell surface-associated aspartic proteases known as yapsins (Yps). While YPS gene deletion mutants have proposed roles in cellular homeostasis, their precise contribution to fungal virulence and host interactions remains unclear. Herein, we present the first detailed biochemical and functional characterization of two native Yps proteins, Yps3 and Yps9, purified from C. glabrata cultures. Both proteases displayed robust activity in a mildly acidic to neutral pH range (5.5-7.0), resistance to the classical aspartic protease inhibitor pepstatin A, and selectively degraded key host antimicrobial peptides, including LL-37 cathelicidin, histatin 5 (Hst5), and kininogen-derived peptide NAT26, by hydrolyzing lysine residues. Additionally, Yps9 promoted C. albicans biofilm dispersal. In a Galleria mellonella infection model, a pre-treatment with each protease enhanced larval survival and increased phenol oxidase activity, implying a role of yapsins in immune priming. Collectively, these findings reveal multifunctional roles for Yps3 and Yps9 in fungal virulence, biofilm modulation, and host immune interactions.
光滑念珠菌（目前归类为光滑纳卡酵母）是一种机会性酵母样真菌，可导致人类感染，由于对抗真菌药物耐药，治疗选择有限。与产生参与致病性的分泌天冬氨酸蛋白酶（Saps）的白色念珠菌不同，光滑念珠菌表达一组独特的细胞表面相关天冬氨酸蛋白酶，称为yapsins（Yps）。虽然YPS基因缺失突变体在细胞内稳态中具有推测的作用，但其对真菌毒力和宿主相互作用的确切贡献仍不清楚。在此，我们首次对从光滑念珠菌培养物中纯化的两种天然Yps蛋白Yps3和Yps9进行了详细的生化和功能表征。这两种蛋白酶在轻度酸性至中性pH范围（5.5 - 7.0）内均表现出强大的活性，对经典天冬氨酸蛋白酶抑制剂胃蛋白酶抑制剂A具有抗性，并通过水解赖氨酸残基选择性降解关键的宿主抗菌肽，包括LL - 37 抗菌肽、组蛋白5（Hst5）和激肽原衍生肽NAT26。此外，Yps9促进白色念珠菌生物膜的分散。在大蜡螟感染模型中，用每种蛋白酶预处理可提高幼虫存活率并增加酚氧化酶活性，这意味着yapsins在免疫启动中发挥作用。总的来说，这些发现揭示了Yps3和Yps9在真菌毒力、生物膜调节和宿主免疫相互作用中的多功能作用。
