Adhesin Amyloid Biology

粘附素淀粉样蛋白生物学

基本信息

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
项目类别：
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

项目摘要

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；但是，允许或限制淀粉样蛋白交叉的分子和结构决定因素种子很难定义。达到这个目标，确定淀粉样蛋白相互作用将是有益的更大的规模，产生一个知识基础，该知识基础可以有助于识别有关跨的规则淀粉样蛋白之间的播种。粘附素是真菌生物学中心的功能性淀粉样蛋白的重要家族和毒力。粘合剂是细胞壁连接的蛋白质，可介导真菌丝状细胞细胞粘附生长和生物膜形成。值得注意的是，近90％的粘合剂是预测的功能性淀粉样蛋白，许多粘合剂已被证明具有能够在溶液中形成淀粉样蛋白原纤维的核心序列。在酵母菌S. cerevisiae和C. belicans，粘附蛋白聚集以形成细胞表面纳米域斑块很重要在增强的伪植物和菌丝丝的细胞细胞粘附中。在之间过渡的能力在机会性人类真菌病原体中毒力需要酵母菌状和丝状生长形式。白色疾病和粘附素已被鉴定为毒力因子。具有强的粘合素序列野生型细胞 - 细胞粘附和细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合剂介导的，我们假设淀粉样相互作用的特异性是亲属歧视和细胞自我认识的基础酵母。我们的初步数据与该假设一致，确定了重要的序列变化在丝状菌株和非丝状菌株之间良好研究的粘附素Flo11p的淀粉样蛋白生成区域 S. cerevisiae。我们还确定，淀粉样蛋白中的单氨基酸变化可以强烈影响交叉播种特异性。在这里，我们建议从丝状和非 - 酿酒酵母的丝状菌株，用于交叉种子。将测试粘附素相互作用的物种障碍使用酿酒酵母和白色念珠菌的一组鉴定的粘合剂。粘合剂具有丰富的信息调查淀粉样相互作用特异性的平台和全基因组功能的平台淀粉样蛋白将使用粘附素框架作为淀粉样蛋白生成潜力和异型相互作用进行测试一个新颖的筛选脚手架。总的来说，这项工作将确定限制的特异性和障碍酵母中的异型粘附素相互作用，同时对淀粉样蛋白交叉种子规则提供更广泛的见解。使用粘合剂作为酵母表面展示的脚手架，我们提出了一种适合高通量的方法鉴定新淀粉样蛋白的应用和特定聚集障碍的快速评估。粘附素是真菌特异性的毒力决定因素与新抗真菌治疗的靶标相关的。