Involvement of N- and C-terminal region of recombinant cervid prion protein in its reactivity to CWD and atypical BSE prions in real-time quaking-induced conversion reaction in the presence of high concentrations of tissue homogenates.
Involvement of N- and C-terminal region of recombinant cervid prion protein in its reactivity to CWD and atypical BSE prions in real-time quaking-induced conversion reaction in the presence of high concentrations of tissue homogenates.
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DOI：
10.1080/19336896.2020.1858694
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发表时间：
2020-12
期刊：
影响因子：
2.3
通讯作者：
Horiuchi M
中科院分区：
文献类型：
--
作者：
Suzuki A;Sawada K;Yamasaki T;Denkers ND;Mathiason CK;Hoover EA;Horiuchi M
实时抖动诱导转换(RT-QuIC)反应是一种灵敏而特异的检测病毒的方法。然而，组织匀浆中存在的抑制因子很容易干扰这一反应。为了确定在高浓度脑组织匀浆存在的情况下，可以检测到低水平的慢性消耗性疾病(CWD)和牛海绵状脑病(BSE)病毒的RT-QuIC条件，我们测试了各种重组病毒蛋白(RPrPs)的反应性。在所测试的rPrP中，重组Cervid PrP(RCerPrP)显示出独特的反应性：rCerPrP对CWD和非典型BSE PrP的反应性不受高浓度正常脑匀浆的高度影响。当N-末端区域(aa25-93)被截断时，rCerPrP的独特反应性消失。用CerPrP的相应区域替换小鼠(Mo)PrP的23-149位氨基酸，部分恢复了rCerPrP在RT-QuIC中的独特反应性。将MoPrP AA 219-231的极端C-末端区域替换为CerPrP的相应区域，部分地授予了rCerPrP对rMoPrP的独特反应活性，表明N-末端和C-末端都参与了rMoPrP。此外，rCerN-Mo-CerCPrP是由CerPrP AA 25-153、MoPrP AA 150-218和CerPrP AA 223-233组成的嵌合PrP，表现出N-末端和C-末端区域的相加效应。这些结果为利用rCerPrP检测CWD和非典型疯牛病病毒提供了机制上的启示，并对RT-QuIC的进一步改进有帮助。
The real-time quaking-induced conversion (RT-QuIC) reaction is a sensitive and specific method for detecting prions. However, inhibitory factors present in tissue homogenates can easily interfere with this reaction. To identify the RT-QuIC condition under which low levels of chronic wasting disease (CWD) and bovine spongiform encephalopathy (BSE) prions can be detected in the presence of high concentrations of brain tissue homogenates, reactivities of various recombinant prion proteins (rPrPs) were tested. Among the tested rPrPs, recombinant cervid PrP (rCerPrP) showed a unique reactivity: the reactivity of rCerPrP to CWD and atypical BSE prions was not highly affected by high concentrations of normal brain homogenates. The unique reactivity of rCerPrP disappeared when the N-terminal region (aa 25–93) was truncated. Replacement of aa 23–149 of mouse (Mo) PrP with the corresponding region of CerPrP partially restored the unique reactivity of rCerPrP in RT-QuIC. Replacement of the extreme C-terminal region of MoPrP aa 219–231 to the corresponding region of CerPrP partially conferred the unique reactivity of rCerPrP to rMoPrP, suggesting the involvement of both N- and C-terminal regions. Additionally, rCerN–Mo–CerCPrP, a chimeric PrP comprising CerPrP aa 25–153, MoPrP aa 150–218, and CerPrP aa 223–233, showed an additive effect of the N- and C-terminal regions. These results provide a mechanistic implication for detecting CWD and atypical BSE prions using rCerPrP and are useful for further improvements of RT-QuIC.
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影响因子：
11.8
作者：
Beringue, Vincent;Herzog, Laetitia;Laude, Hubert
通讯作者：
Laude, Hubert
DOI：
10.1073/pnas.0409008102
发表时间：
2005-01-18
影响因子：
11.1
作者：
Gossert, AD;Bonjour, S;Wüthrich, K
通讯作者：
Wüthrich, K
影响因子：
2.9
作者：
Liemann, S;Glockshuber, R
通讯作者：
Glockshuber, R
影响因子：
6.7
作者：
Capobianco, Raffaella;Casalone, Cristina;Tagliavini, Fabrizio
通讯作者：
Tagliavini, Fabrizio
影响因子：
48
作者：
Atarashi, Ryuichiro;Moore, Roger A.;Caughey, Byron
通讯作者：
Caughey, Byron