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Candidacidal mechanism of the arenicin-3-derived peptide NZ17074 from Arenicola marina

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Abstract

The candidacidal mechanisms of NZ17074, which is a variant of arenicin-3 from Arenicola marina, against human pathogenic fungus Candida albicans are reported in this work. The minimum inhibitory concentration (MIC) of NZ17074 toward C. albicans was 4 μg/ml, and this peptide exerted marked candidacidal activity in an energy-dependent and salt-sensitive manner. The flow cytometric analysis using propidium iodide (PI) showed that the plasma membrane of cells treated with NZ17074 was perturbed and that the cells were arrested in the G2/M phase. The dihydrorhodamine-123 (DHR-123) staining showed that the reactive oxygen species (ROS) production of C. albicans increased after exposure to NZ17074. Typical cellular disruption events, such as mitochondrial degradation, nuclear fragmentation, nuclear membrane disruption, and chromatin condensation, were further revealed through rhodamine 123 (RH123) staining, 4′,6-diamidino-2-phenylindole (DAPI) staining, and transmission electron microscopy. In addition, the intracellular localization of this peptide was concentration dependent: it was located in the membrane at low concentrations (4 to 8 μg/ml) and penetrated into the cytoplasm at high concentrations (16 to 32 μg/ml). Our results suggested that NZ17074 exerts its candidacidal effects by disrupting the cell membrane, inducing apoptosis, and interrupting the cell cycle. These findings showed the potential of NZ17074 as a new candidacidal peptide, in addition to its antibacterial activities.

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Acknowledgments

The authors wish to acknowledge Tong Zhao and Jingnan Liang from the Core Facility at the Institute of Microbiology at the Chinese Academy of Sciences (CAS) for their technical support with the flow cytometric and TEM analyses and Jing Wu from the Institute of Genetics and Developmental Biology at CAS for her coordination of the confocal fluorescence microscopy analysis. This study was supported by the National Natural Science Foundation of China (No. 31372346, No. 31302004, and No. 30972125), the Project of the National Support Program for Science and Technology in China (No. 2013BAD10B02 and No. 2011BAD26B02), and the AMP Direction of Innovation Program of Agric Sci & Tech in CAAS (2013–2017).

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Authors and Affiliations

  1. Key Laboratory of Feed Biotechnology, Ministry of Agriculture, Beijing, 100081, China

    Xiaojie Wang, Xiumin Wang, Da Teng, Yong Zhang, Ruoyu Mao, Di Xi & Jianhua Wang

  2. Gene Engineering Laboratory, Feed Research Institute, Chinese Academy of Agricultural Sciences, 12 Zhongguancun Nandajie St., Haidian District, Beijing, 100081, People’s Republic of China

    Xiaojie Wang, Xiumin Wang, Da Teng, Yong Zhang, Ruoyu Mao, Di Xi & Jianhua Wang

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  1. Xiaojie Wang
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Correspondence to Jianhua Wang.

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Xiaojie Wang and Xiumin Wang contributed equally to this paper.

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Wang, X., Wang, X., Teng, D. et al. Candidacidal mechanism of the arenicin-3-derived peptide NZ17074 from Arenicola marina . Appl Microbiol Biotechnol 98, 7387–7398 (2014). https://doi.org/10.1007/s00253-014-5784-6

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