Adhesin Amyloid Biology

粘附素淀粉样蛋白生物学

• 批准号: 10726038
• 负责人: Jason E Gestwicki
• 金额: $ 20.35万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10726038
• 关键词: Adhesions; Affect; Amino Acids; Amyloid; Amyloid Fibrils; Amyloid Proteins; Antifungal Agents; Antifungal Therapy; Bacterial Adhesins; Binding; Biological Assay; Biology; Candida albicans; Cell Adhesion; Cell Communication; Cell Wall; Cell surface; Cell-Cell Adhesion; Cells; Communities; Data; Development; Discrimination; Disease; Exclusion; Family; Filament; Flocculation; Future; Genetic study; Goals; Growth; Human; Infection; Investigation; Knowledge; Link; Mediating; Methods; Microbial Biofilms; Molecular; Mus; Neurodegenerative Disorders; Outcome; Pathogenesis; Phenotype; Pilot Projects; Point Mutation; Prions; Process; Property; Protein Isoforms; Proteins; Rapid screening; Recombinant Proteins; Role; Saccharomyces cerevisiae; Specificity; Structure; Supporting Cell; Surface; System; Tandem Repeat Sequences; Testing; Variant; Virulence; Virulence Factors; Work; Yeasts; amyloid structure; cell type; drug development; experience; experimental study; fascinate; functional genomics; fungus; genome-wide; glycosylation; high throughput screening; improved; innovation; insight; knowledgebase; microbial; microbial community; nano; novel; pathogenic fungus; protein aggregation; remediation; scaffold; screening; self assembly; structural determinants; tau Proteins

Heterotypic interactions between amyloid proteins are critical in understanding the outcomes of amyloid-based infection and treatment; however, the molecular and structural determinants that allow or limit amyloid cross- seeding have been difficult to define. Towards this goal, it will be informative to identify amyloid interactions on a larger scale, generating a knowledgebase that can contribute to the identification of rules governing cross- seeding among amyloids. The adhesins are an important family of functional amyloids central to fungal biology and virulence. Adhesins are cell wall-attached proteins that mediate cell-cell adhesion in fungal filamentous growth and biofilm formation. Notably, nearly 90% of adhesins are predicted functional amyloids, and many have been demonstrated to possess core sequences capable of forming amyloid fibrils in solution. In the yeasts S. cerevisiae and C. albicans, adhesins aggregate to form cell surface nanodomain patches important in the enhanced cell-cell adhesion of pseudohyphal and hyphal filaments. The ability to transition between yeast-like and filamentous growth forms is required for virulence in the opportunistic human fungal pathogen C. albicans, and adhesins have been identified as virulence factors. Adhesin sequences with strong amyloidogenic potential are required for wild-type cell-cell adhesion and filamentation. Fungi preferentially bind to like cells in biofilms and filamentous communities. Since this binding is mediated through adhesins, we hypothesize that specificity in amyloid interactions underlies kin discrimination and cellular self-recognition in yeast. Our preliminary data are consistent with this hypothesis, identifying important sequence variation in amyloidogenic regions of the well-studied adhesin Flo11p between filamentous and non-filamentous strains of S. cerevisiae. We also identify that single amino acid changes in amyloid proteins can strongly affect cross- seeding specificity. Here, we propose to test adhesin protein isoforms of Flo11p from filamentous and non- filamentous strains of S. cerevisiae for cross-seeding. Species barriers to adhesin interactions will be tested using the set of identified adhesins from S. cerevisiae and C. albicans. The adhesins present an informative platform for the investigation of amyloid interaction specificity, and a genome-wide set of yeast functional amyloids will be tested for amyloidogenic potential and heterotypic interactions using the adhesin framework as a novel screening scaffold. Collectively, this work will determine the specificity and barriers that constrain heterotypic adhesin interactions in yeast, while offering broader insight into the rules of amyloid cross-seeding. Using the adhesins as a scaffold for yeast surface display, we present a method amenable to high throughput applications for the identification of new amyloids and the rapid assessment of specific aggregation barriers. Adhesins are fungal-specific virulence determinants relevant as targets for new antifungal treatments.

淀粉样蛋白之间的异型相互作用对于理解基于淀粉样蛋白的结果至关重要 感染和治疗；然而，允许或限制淀粉样蛋白交叉的分子和结构决定因素 播种很难定义。为了实现这一目标，识别淀粉样蛋白的相互作用将提供丰富的信息。 规模更大，生成一个知识库，有助于识别管理跨领域的规则 淀粉样蛋白之间播种。粘附素是真菌生物学核心的功能性淀粉样蛋白的重要家族 和毒力。粘附素是细胞壁附着的蛋白质，介导真菌丝状细胞中的细胞间粘附 生长和生物膜形成。值得注意的是，近 90% 的粘附素被预测为功能性淀粉样蛋白，并且许多 已被证明拥有能够在溶液中形成淀粉样原纤维的核心序列。在 酿酒酵母和白色念珠菌，粘附素聚集形成细胞表面纳米结构域斑块很重要 增强假菌丝和菌丝细丝的细胞间粘附。之间转换的能力 机会性人类真菌病原体 C. 的毒力需要酵母样和丝状生长形式。 白色念珠菌和粘附素已被确定为毒力因子。粘附素序列具有强 野生型细胞-细胞粘附和成丝需要淀粉样蛋白生成潜力。真菌优先结合 喜欢生物膜和丝状群落中的细胞。由于这种结合是通过粘附素介导的，我们 假设淀粉样蛋白相互作用的特异性是亲属歧视和细胞自我识别的基础 酵母。我们的初步数据与这一假设一致，确定了重要的序列变异 丝状和非丝状菌株之间经过充分研究的粘附素 Flo11p 的淀粉样蛋白生成区域 酿酒酵母。我们还发现淀粉样蛋白中的单个氨基酸变化可以强烈影响交叉 播种特异性。在这里，我们建议测试丝状和非丝状 Flo11p 的粘附素蛋白亚型。 用于交叉播种的酿酒酵母丝状菌株。将测试粘附素相互作用的物种障碍 使用来自酿酒酵母和白色念珠菌的一组已鉴定的粘附素。粘附素提供了丰富的信息 用于研究淀粉样蛋白相互作用特异性的平台，以及一组全基因组酵母功能 将使用粘附素框架测试淀粉样蛋白的淀粉样蛋白生成潜力和异型相互作用 一种新颖的筛选支架。总的来说，这项工作将确定限制的特殊性和障碍 酵母中的异型粘附素相互作用，同时提供对淀粉样蛋白交叉播种规则的更广泛的了解。 使用粘附素作为酵母表面展示的支架，我们提出了一种适合高通量的方法 用于鉴定新淀粉样蛋白和快速评估特定聚集障碍的应用。 粘附素是真菌特异性毒力决定因素，可作为新抗真菌治疗的靶标。