Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, New Hampshire, USA.
Proteins. 2022 Mar;90(3):869-880. doi: 10.1002/prot.26282. Epub 2021 Dec 2.
Prions, misfolded proteins that aggregate, cause an array of progressively deteriorating conditions to which, currently, there are no effective treatments. The presently accepted model indicates that the soluble non-prion forms of prion-forming proteins, such as the well-studied SUP35, do not exist in large aggregated molecular complexes. Here, we show using analytical ultracentrifugation with fluorescent detection that the non-prion form of SUP35 exists in a range of discretely sized soluble complexes (19S, 28S, 39S, 57S, and 70S-200S). Similar to the [PSI+] aggregated complexes, each of these [psi-] complexes associates at stoichiometric levels with a large variety of molecular chaperones: HSP70 proteins comprise the major component. Another yeast prion-forming protein, RNQ1 (known to promote the production of the prion SUP35 state), is also present in SUP35 complexes. These results establish that the non-prion SUP35, like its prion form, is predisposed to form large molecular complexes containing chaperones and other prion-forming proteins. These results agree with our previous studies on the huntingtin protein. That the normal forms for aggregation-prone proteins may preexist in large molecular complexes has important ramifications for the progression of diseases involving protein aggregation.
朊病毒是一种错误折叠的蛋白质,会聚集并导致一系列逐渐恶化的病症,而目前尚无有效的治疗方法。目前公认的模型表明,朊病毒形成蛋白的可溶性非朊病毒形式,如研究充分的 SUP35,不存在于大型聚集的分子复合物中。在这里,我们使用带有荧光检测的分析超速离心法表明,SUP35 的非朊病毒形式存在于一系列离散大小的可溶性复合物(19S、28S、39S、57S 和 70S-200S)中。与 [PSI+] 聚集复合物类似,这些 [psi-] 复合物以化学计量水平与各种分子伴侣结合:HSP70 蛋白构成主要成分。另一种酵母朊病毒形成蛋白 RNQ1(已知可促进朊病毒 SUP35 状态的产生)也存在于 SUP35 复合物中。这些结果表明,非朊病毒 SUP35 与其朊病毒形式一样,容易形成包含伴侣蛋白和其他朊病毒形成蛋白的大型分子复合物。这些结果与我们之前对亨廷顿蛋白的研究一致。易于聚集的蛋白质的正常形式可能预先存在于大型分子复合物中,这对涉及蛋白质聚集的疾病的进展具有重要影响。