Stehli Daniel, Mulaj Mentor, Miti Tatiana, Traina Joshua, Foley Joseph, Muschol Martin
Department of Physics; University of South Florida ; Tampa, FL USA.
Intrinsically Disord Proteins. 2015 Jun 10;3(1):e1056905. doi: 10.1080/21690707.2015.1056905. eCollection 2015.
Self-assembly of proteins and peptides into amyloid fibrils involves multiple distinct intermediates and late-stage fibrillar polymorphs. Understanding the conditions and mechanisms that promote the formation of one type of intermediate and polymorph over the other represents a fundamental challenge. Answers to this question are also of immediate biomedical relevance since different amyloid aggregate species have been shown to have distinct pathogenic potencies. One amyloid polymorph that has received comparatively little attention are amyloid spherulites. Here we report that self-assembly of the intrinsically disordered polymer poly(L-glutamic) acid (PLE) can generate amyloid spherulites. We characterize spherulite growth kinetics, as well as the morphological, optical and tinctorial features of this amyloid polymorph previously unreported for PLE. We find that PLE spherulites share both tinctorial and structural characteristics with their amyloid fibril counterparts. Differences in PLE's molecular weight, polydispersity or chemistry could not explain the selective propensity toward either fibril or spherulite formation. Instead, we provide evidence that PLE polymers can exist in either a collapsed globule or an extended random coil conformation. The collapsed globule consistently produces spherulites while the extended coil assembles into disordered fibril bundles. This results suggests that these 2 PLE conformers directly affect the morphology of the resulting macroscopic amyloid assembly.
蛋白质和肽自组装成淀粉样纤维涉及多个不同的中间体和晚期纤维多晶型物。了解促进一种中间体和多晶型物而非另一种形成的条件和机制是一项基本挑战。由于已表明不同的淀粉样聚集体具有不同的致病能力,这个问题的答案也具有直接的生物医学相关性。一种相对较少受到关注的淀粉样多晶型物是淀粉样球晶。在此我们报告,内在无序聚合物聚(L-谷氨酸)(PLE)的自组装可以产生淀粉样球晶。我们表征了球晶的生长动力学,以及这种淀粉样多晶型物的形态、光学和染色特征,这些特征此前未针对PLE进行过报道。我们发现PLE球晶与其淀粉样纤维对应物具有染色和结构特征。PLE的分子量、多分散性或化学性质的差异无法解释其对纤维或球晶形成的选择性倾向。相反,我们提供证据表明PLE聚合物可以以塌陷球状体或伸展无规卷曲构象存在。塌陷球状体始终产生球晶,而伸展卷曲则组装成无序的纤维束。这一结果表明,这两种PLE构象直接影响最终宏观淀粉样聚集体的形态。