Development and structural determination of an anti-PrPC aptamer that blocks pathological conformational conversion of prion protein
Development and structural determination of an anti-PrPC aptamer that blocks pathological conformational conversion of prion protein
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阻断朊病毒蛋白病理构象转变的抗 PrPC 适体的开发和结构测定
DOI：
10.1038/s41598-020-61966-4
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发表时间：
2020
影响因子：
4.6
通讯作者：
Katahira Masato
中科院分区：
文献类型：
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作者：
Mashima Tsukasa;Lee Joon-Hwa;Kamatari Yuji O.;Hayashi Tomohiko;Nagata Takashi;Nishikawa Fumiko;Nishikawa Satoshi;Kinoshita Masahiro;Kuwata Kazuo;Katahira Masato
朊病毒疾病包括由朊病毒蛋白从细胞（PrPC）转化为病理（PrPSc）同种型引起的致命性神经病。之前，我们获得了RNA适体r（GGAGGAGGAGGA）（R12），其折叠成独特的G-四链体。R12同二聚体与PrPC分子结合，抑制PrPC转化为PrPSc。在这里，我们开发了一种新的RNA适体r（GGAGGAGGAGGAGGAGGAGGA）（R24），其中两个R12串联连接。R24对羊瘙痒病感染细胞系中PrPSc形成的半数抑制浓度（IC_（50））约为0.001 μ g/ml。100 nM，即，比R12低两个数量级。除部分抗体外，R24的IC_（50）最低，抗朊病毒活性最高。我们还开发了一种相关的适体r（GGAGGAGGAGGA-A-GGAGGAGGAGGA）（R12-A-R12），IC 50约为100。500 nM。通过NMR确定的单个R12-A-R12分子的结构类似于R12同二聚体的结构。R24或R12-A-R12的四链体结构是单分子的，因此当它们被给予朊病毒感染的细胞培养物时，该结构可以稳定地形成。这可能是它们能够发挥高抗朊病毒活性的原因。
Prion diseases comprise a fatal neuropathy caused by the conversion of prion protein from a cellular (PrPC) to a pathological (PrPSc) isoform. Previously, we obtained an RNA aptamer, r(GGAGGAGGAGGA) (R12), that folds into a unique G-quadruplex. The R12 homodimer binds to a PrPCmolecule, inhibiting PrPC-to-PrPScconversion. Here, we developed a new RNA aptamer, r(GGAGGAGGAGGAGGAGGAGGAGGA) (R24), where two R12s are tandemly connected. The 50% inhibitory concentration for the formation of PrPSc(IC50) of R24 in scrapie-infected cell lines was ca. 100 nM, i.e., much lower than that of R12 by two orders. Except for some antibodies, R24 exhibited the lowest recorded IC50and the highest anti-prion activity. We also developed a related aptamer, r(GGAGGAGGAGGA-A-GGAGGAGGAGGA) (R12-A-R12), IC50being ca. 500 nM. The structure of a single R12-A-R12 molecule determined by NMR resembled that of the R12 homodimer. The quadruplex structure of either R24 or R12-A-R12 is unimolecular, and therefore the structure could be stably formed when they are administered to a prion-infected cell culture. This may be the reason they can exert high anti-prion activity.