Zdybicka-Barabas Agnieszka, Stączek Sylwia, Mak Paweł, Kapral-Piotrowska Justyna, Skrzypiec Krzysztof, Wydrych Jerzy, Pawlikowska-Pawlęga Bożena, Gruszecki Wiesław I, Cytryńska Małgorzata
Department of Immunobiology, Institute of Biological Sciences, Faculty of Biology and Biotechnology, Maria Curie-Skłodowska University, Akademicka 19 St., 20-033 Lublin, Poland.
Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, Gronostajowa 7 St., 30-387 Krakow, Poland.
Int J Mol Sci. 2025 Aug 29;26(17):8438. doi: 10.3390/ijms26178438.
Two proline-rich antimicrobial peptides (PrAMPs), named P1 and P2, purified from hemolymph of the greater wax moth , were studied for their effects on Gram-negative () and Gram-positive () bacteria. Both peptides decreased the bacterial survival rate and caused bacterial membrane permeabilization. However, in both cases, the P2 peptide was approximately three times more effective than the P1 peptide. Fluorescence microscopy imaging demonstrated binding of both FITC-labeled peptides to and cells. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) imaging of peptide-treated bacteria revealed considerable changes in cell morphology, cell surface topography, and nanomechanical properties. The interactions of the PrAMPs with bacterial cells were also analyzed by FTIR spectroscopy. The P1 peptide action toward led to partial aggregation of proteins, whereas treatment with P2 resulted in reduced protein aggregation, reflecting differences between both PrAMPs antibacterial action. Moreover, both PrAMPs caused a decrease and an increase in the protein content in relation to lipids on the and cell surface, respectively. The obtained results reflect not only differences between the P1 and P2 peptides but also differences in the cell surface between Gram-negative and Gram-positive bacteria. Both characterized PrAMPs are further representatives of proline-rich peptides with a membrane-permeabilizing antimicrobial mode of action.
从大蜡螟血淋巴中纯化得到两种富含脯氨酸的抗菌肽（PrAMPs），分别命名为P1和P2，并研究了它们对革兰氏阴性菌（）和革兰氏阳性菌（）的作用。两种肽均降低了细菌存活率并导致细菌细胞膜通透性增加。然而，在这两种情况下，P2肽的效果大约是P1肽的三倍。荧光显微镜成像显示两种异硫氰酸荧光素（FITC）标记的肽均能与和细胞结合。对经肽处理的细菌进行原子力显微镜（AFM）和扫描电子显微镜（SEM）成像，结果显示细胞形态、细胞表面形貌和纳米力学性能发生了显著变化。还通过傅里叶变换红外光谱（FTIR）分析了PrAMPs与细菌细胞的相互作用。P1肽对的作用导致蛋白质部分聚集，而用P2处理则导致蛋白质聚集减少，这反映了两种PrAMPs抗菌作用的差异。此外，两种PrAMPs分别导致革兰氏阴性菌和革兰氏阳性菌细胞表面蛋白质含量相对于脂质含量降低和增加。所得结果不仅反映了P1和P2肽之间的差异，也反映了革兰氏阴性菌和革兰氏阳性菌细胞表面的差异。这两种已表征的PrAMPs是具有膜通透性抗菌作用模式的富含脯氨酸肽的进一步代表。
