Structures of non-prion amyloids

非朊病毒淀粉样蛋白的结构

• 批准号: 8349933
• 负责人: Reed B. WICKNER
• 金额: $ 29.3万
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8349933
• 关键词: Acids; Adhesions; Amyloid; Amyloid Fibrils; Amyloidosis; Amyotrophic Lateral Sclerosis; Anabolism; Architecture; Biogenesis; Cell Adhesion; Cell surface; Cells; Collaborations; Complement; Cytoplasm; Escherichia coli; Escherichia coli Proteins; Filament; Growth; Homologous Gene; Human; Melanins; Melanosomes; Mus; Neurons; Organism; Orthologous Gene; Patients; Prealbumin; Prion Diseases; Proteins; RNA-Binding Protein FUS; Role; Serum Proteins; Structure; Yeast Model System; Yeasts; Zebrafish; aging population; amyloid formation; amyloid structure; base; beta pleated sheet; cell growth; melanocyte; non-prion; pMel17 antigen; protein TDP-43; yeast prion

We have studied several non-prion amyloids that have normal functions for their respective organisms: Pmel17, a mammalian melanocyte protein that normally forms amyloid involved in melanin biosynthesis and curli, a cell-surface amyloid of E. coli that is involved in adhesion. We found that the repeat domain of human Pmel17 is able to form an amyloid structure that forms readily at the mildly acid pH typical of the melanocyte, and rapidly dissolves at neutral pH (1). The repeat domains of Pmel17 homologs from mouse and zebrafish have little conservation of sequence, but do conserve the ability to form amyloid specifically at the acid pH of the melanosome (2). We characterized the amyloid form by curli proteins and found that they are not in the in-register parallel architecture typical of the yeast prion amyloids (3). In collaboration with the Frank Shewmaker (USUHS), we also studied the FUS protein that is found aggregated in the cytoplasm of neurons of patients with amyotrophic lateral sclerosis. We found that FUS aggregates in yeast cells, producing a marked growth inhibition (4). As in ALS neurons, FUS tends to co-aggregate with TDP-43 when co-expressed in yeast (4). 1. McGlinchey RP, Shewmaker F, McPhie P, Monterroso B, Thurber KR & Wickner RB (2009) The repeat domain of the melanosome fibril protein Pmel17 forms the amyloid core promoting melanin synthesis. Proc. Natl. Acad. Sci. USA 106: 13731-6. 2. McGlinchey, R., Shewmaker, F., Hu, K. H., McPhie, P., Tycko, R., Wickner, RB (2011) Repeat domains of melanosome matrix protein Pmel17 orthologs form amyloid fibrils at the acidic melanosomal pH. J. Biol. Chem. 286: 8385-93. 3. Shewmaker F, McGlinchey R, Thurber KR, McPhie P, Dyda F, Tycko R & Wickner RB (2009) The functional curli amyloid is not based on in-register parallel beta-sheet structure. J. Biol. Chem. 284: 25065 - 25076. 4. Kryndushkin DS, Wickner RB, Shewmaker F. FUS/TLS forms cytoplasmic aggregates, inhibits cell growth and interacts with TDP-43 in a yeast model of amyotrophic lateral sclerosis. Protein Cell 2011; 2:223 - 36.

我们研究了几种非蛋白质淀粉样蛋白具有正常功能的非蛋白淀粉样蛋白：PMEL17，一种通常形成与黑色素生物合成的淀粉样蛋白的哺乳动物黑色素细胞蛋白和涉及大肠杆菌的细胞表面淀粉样蛋白酶的淀粉样蛋白。 我们发现，人PMEL17的重复结构域能够形成一种淀粉样结构，该结构很容易在黑素细胞的典型酸中pH下形成，并在中性pH下迅速溶解（1）。 来自小鼠和斑马鱼的PMEL17同源物的重复结构域几乎没有序列的保守性，但确实保留了在黑色素体的酸pH值下形成淀粉样蛋白的能力（2）。 我们通过Curli蛋白表征了淀粉样蛋白形式，发现它们不在酵母菌prion蛋白淀粉样蛋白的典型内注册平行结构中（3）。 在与弗兰克·谢玛克（Frank Shewmaker）（USUHS）的合作下，我们还研究了肌萎缩性侧面硬化症患者神经元中骨质骨质中发现的FUS蛋白。 我们发现酵母细胞中的FUS聚集体产生明显的生长抑制作用（4）。 与ALS神经元一样，当在酵母中共表达时，FUS倾向于与TDP-43共聚集（4）。 1。McGlincheyRP，Shewmaker F，McPhie P，Monterroso B，Thurber KR＆Wickner RB（2009）黑色素纤维蛋白PMEL17的重复结构域形成促进黑色素合成的淀粉样蛋白核心。 Proc。纳特。学院。科学。美国106：13731-6。 2。McGlinchey，R。，Shewmaker，F.，Hu，K。H.，McPhie，P.，Tycko，R.，Wickner，RB（2011）黑色素体基质蛋白PMEL17直系同源物的重复结构域形成酸性黑色素体pH的淀粉样纤维。 J. Biol。化学286：8385-93。 3。ShewmakerF，McGlinchey R，Thurber KR，McPhie P，Dyda F，Tycko R＆Wickner RB（2009）功能性的CURLI淀粉样蛋白不是基于注册的平行β-表 - β-表 - β-图。 J. Biol。化学284：25065-25076。 4。KryndushkinDS，Wickner RB，Shewmaker F. FUS/TLS形成细胞质聚集体，抑制细胞的生长并在肌营养性侧面硬化症的酵母模型中与TDP-43相互作用。蛋白质细胞2011; 2：223-36。