Perforin-2 clockwise hand-over-hand pre-pore to pore transition mechanism.
Perforin-2 clockwise hand-over-hand pre-pore to pore transition mechanism.
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DOI：
10.1038/s41467-022-32757-4
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发表时间：
2022-08-26
影响因子：
16.6
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中科院分区：
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--
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穿孔素-2(PFN2，MPEG1)是一种成孔蛋白，作为哺乳动物免疫系统的第一道防线，迅速杀死巨噬细胞吞噬溶酶体中被吞噬的微生物。PFN2自组装成十六聚体前孔环，在酸化后过渡到破坏靶细胞膜的孔中。在这里，我们利用高速原子力显微镜(HS-AFM)成像、行扫描和分子动力学模拟，阐明了PFN_2的预孔-孔转变途径和动力学。酸化后，前孔环(前孔-I)频繁出现，1.8 S−1，开环动力学，最终，0.2 S−1，开始过渡到中间的、短暂的、~75 ms的前孔-II状态，诱导顺时针方向的前孔-I-前孔-II扩展。相应地，第一个前孔II亚基经历了主要的构象变化，孔态也以~15 S−1的速率顺时针传播。因此，预孔到孔的转变是一种顺时针交接机制，是在~1.3MCPF/CDC超家族中完成的。我们的发现表明，膜的顺时针插入机制可能是通用的，随着变化的变化。哺乳动物免疫防御复合体穿孔素-2(PFN2)从前孔到孔的结构转换的直接可视化揭示了一种顺时针交接机制，该机制以每秒~15个亚基的速度传播。
Perforin-2 (PFN2, MPEG1) is a pore-forming protein that acts as a first line of defense in the mammalian immune system, rapidly killing engulfed microbes within the phagolysosome in macrophages. PFN2 self-assembles into hexadecameric pre-pore rings that transition upon acidification into pores damaging target cell membranes. Here, using high-speed atomic force microscopy (HS-AFM) imaging and line-scanning and molecular dynamics simulation, we elucidate PFN2 pre-pore to pore transition pathways and dynamics. Upon acidification, the pre-pore rings (pre-pore-I) display frequent, 1.8 s−1, ring-opening dynamics that eventually, 0.2 s−1, initiate transition into an intermediate, short-lived, ~75 ms, pre-pore-II state, inducing a clockwise pre-pore-I to pre-pore-II propagation. Concomitantly, the first pre-pore-II subunit, undergoes a major conformational change to the pore state that propagates also clockwise at a rate ~15 s−1. Thus, the pre-pore to pore transition is a clockwise hand-over-hand mechanism that is accomplished within ~1.3 s. Our findings suggest a clockwise mechanism of membrane insertion that with variations may be general for the MACPF/CDC superfamily. Direct visualization of the structural transformation from pre-pore to pore of the mammalian immune defense complex perforin-2 (PFN2) reveals a clockwise hand-over-hand mechanism that propagates at ~15 subunits per second.
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