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α-螺旋肽的高两亲性增强了未甲基化 CpG DNA 对鼠巨噬样 RAW264.7 细胞的激活作用。

High amphipathicity of α-helical peptides enhances unmethylated CpG DNA-induced activation of mouse macrophage-like RAW264.7 cells.

机构信息

Faculty of Pharmaceutical Sciences, Doshisha Women's College of Liberal Arts, Kyotanabe, Kyoto, 610-0395, Japan.

出版信息

Sci Rep. 2024 Jul 15;14(1):16274. doi: 10.1038/s41598-024-67166-8.

DOI:10.1038/s41598-024-67166-8
PMID:39009614
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11251158/
Abstract

The α-helical antimicrobial peptide Kn2-7 enhances the activation of mouse macrophage-like RAW264.7 induced by DNA containing unmethylated cytosine-guanine motifs (CpG DNA). This enhancement is related to increased cellular uptake of DNA by Kn2-7, but the relevant properties of Kn2-7 are unknown. Physicochemical property analysis revealed that Kn2-7 has high amphipathicity. In contrast, the α-helical antimicrobial peptide L5, which increases the cellular uptake of CpG DNA but does not enhance CpG DNA-induced activation, has low amphipathicity. Kn2-7 derivatives with decreased amphipathicity but the same amino acid composition as Kn2-7 did not enhance CpG DNA-induced activation. On the other hand, L5 derivatives with high amphipathicity but the same amino acid composition as L5 enhanced CpG DNA-induced activation. Cellular uptake of DNA was not increased by the L5 derivatives, indicating that high amphipathicity does not affect DNA uptake. Furthermore, α-helical peptides with reversed sequences relative to the Kn2-7 and L5 derivatives with high amphipathicity were synthesized. The reversed-sequence peptides, which had the same amphipathicity but different amino acid sequences from their counterparts, enhanced CpG DNA-induced activation. Taken together, these observations indicate that the high amphipathicity of α-helical peptides enhances the CpG DNA-induced activation of RAW264.7.

摘要

α-螺旋抗菌肽 Kn2-7 增强了含有未甲基化胞嘧啶-鸟嘌呤基序（CpG DNA）的 DNA 对小鼠巨噬细胞样 RAW264.7 的激活。这种增强与 Kn2-7 增加 DNA 的细胞摄取有关，但 Kn2-7 的相关性质尚不清楚。物理化学性质分析表明 Kn2-7 具有高两亲性。相比之下，增加 CpG DNA 细胞摄取但不增强 CpG DNA 诱导的激活的 α-螺旋抗菌肽 L5 具有低两亲性。具有降低的两亲性但与 Kn2-7 相同的氨基酸组成的 Kn2-7 衍生物不能增强 CpG DNA 诱导的激活。另一方面，具有高两亲性但与 L5 相同的氨基酸组成的 L5 衍生物增强了 CpG DNA 诱导的激活。L5 衍生物并未增加 DNA 的摄取，表明高两亲性不会影响 DNA 的摄取。此外，还合成了相对于 Kn2-7 具有反转序列的 α-螺旋肽和具有高两亲性的 L5 衍生物。与对照肽具有相同两亲性但氨基酸序列不同的反转序列肽增强了 CpG DNA 诱导的激活。综上所述，这些观察结果表明 α-螺旋肽的高两亲性增强了 CpG DNA 诱导的 RAW264.7 激活。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/98de0fc3e23c/41598_2024_67166_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/77d99de585ab/41598_2024_67166_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/afc00009d885/41598_2024_67166_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/625471c1a399/41598_2024_67166_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/67d4edeaca07/41598_2024_67166_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/da0edf8eaa60/41598_2024_67166_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/6022e7651f54/41598_2024_67166_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/98de0fc3e23c/41598_2024_67166_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/77d99de585ab/41598_2024_67166_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/afc00009d885/41598_2024_67166_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/625471c1a399/41598_2024_67166_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/67d4edeaca07/41598_2024_67166_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/da0edf8eaa60/41598_2024_67166_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/6022e7651f54/41598_2024_67166_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8c0/11251158/98de0fc3e23c/41598_2024_67166_Fig7_HTML.jpg

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本文引用的文献

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Enhanced cellular uptake of CpG DNA by α-helical antimicrobial peptide Kn2-7: Effects on macrophage responsiveness to CpG DNA.
α-螺旋抗菌肽 Kn2-7增强 CpG DNA 的细胞摄取：对巨噬细胞对 CpG DNA 反应性的影响。
Biochem Biophys Res Commun. 2020 Sep 10;530(1):100-106. doi: 10.1016/j.bbrc.2020.07.013. Epub 2020 Jul 30.
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