Cell Surface Receptor Recognition and Membrane Fusion in Mammalian Fertilization

哺乳动物受精中的细胞表面受体识别和膜融合

• 批准号: 10396577
• 负责人: Shaogeng Tang
• 金额: $ 12.75万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10396577
• 关键词: Adhesions; Advisory Committees; Affinity; Architecture; Bacterial Adhesins; Binding; Biochemistry; Biological; Biological Assay; Biological Process; CRISPR-mediated transcriptional activation; Candidate Disease Gene; Cell Surface Proteins; Cell Surface Receptors; Cell fusion; Cell membrane; Cell surface; Cells; Cellular biology; Characteristics; Charge; Coiled-Coil Domain; Complementary DNA; Cultured Cells; Diploidy; Disulfides; ERp57; Electronic Medical Records and Genomics Network; Electrostatics; Environment; Enzymatic Biochemistry; Event; Exhibits; Fertility; Fertilization; Fluorescence-Activated Cell Sorting; Genetic; Genetic Screening; Genetic screening method; Genetic study; Germ Cells; Goals; Haploidy; Higher Order Chromatin Structure; Human; Human Engineering; Immunoglobulins; In Vitro; Intercellular Junctions; Laboratories; Libraries; Life; Ligands; Liposomes; Mediating; Membrane; Membrane Fusion; Membrane Proteins; Mentors; Methods; Molecular; Molecular Conformation; Mus; Oocytes; Organism; Pathway interactions; Phase; Process; Protein Disulfide Isomerase; Protein Engineering; Protein Family; Proteins; Regulation; Reproductive Health; Research; Resolution; Role; Schools; Structure; Sulfhydryl Compounds; Surface; Testing; Training; Universities; Viral; Work; Yeasts; career development; design; egg; extracellular; functional genomics; in vivo; insight; interdisciplinary approach; male; male fertility; mutation screening; novel; paralogous gene; programs; receptor; reconstitution; research and development; self assembly; skills; sperm cell; structural biology

Project Summary/Abstract Fertilization is an essential biological process that involves cell surface recognition, adhesion, and fusion of haploid sperm and egg to form a new, genetically distinct diploid organism. Surprisingly little is known about how plasma membranes interact and fuse, largely due to the limited availability of mammalian gametes and the technical challenges of capturing transient extracellular interactions. While the gamete membrane fusion machinery remains elusive, it is known that a human immunoglobulin superfamily protein, Izumo1, localizes to the equatorial segment of spermatozoon, where it interacts with an oocyte ligand, Juno, before gamete fusion takes place. To date, Izumo1- Juno is the only essential receptor-ligand pair identified in the pathway. Recent genetic studies in mice have identified additional sperm surface proteins essential for male fertility, but their interactions and regulation have not been defined. To close these major gaps, I have designed a research program around the central hypothesis that gamete surface proteins orchestrate their structural rearrangements and membrane remodeling, leading to gamete fusion. I will test my hypothesis by (1) determining the molecular mechanisms of human Izumo1 self-assembly, (2) characterizing the roles of human Izumo family proteins in membrane adhesion and fusion, and (3) identifying additional receptor-ligand interactions essential for gamete fusion and mammalian fertilization. My multidisciplinary approach leverages high- resolution structural analyses to interrogate the functional organization of known essential gamete surface proteins, and harnesses high-throughput forward genetic screens to explore the interacting networks of additional emerging molecules. Crucially, understanding these interactions will elucidate mechanisms of cell-cell recognition and membrane fusion and reveal essential factors that maintain human fertility and reproductive health. From graduate school, I have extensive training in genetics, cell biology, and membrane biochemistry. During my early postdoctoral work, I honed my skills in structural biology of cell surface receptors and protein engineering of viral membrane fusion machines. I will exploit the excellent research environments in the laboratories of my mentor Dr. Peter Kim and my co-mentor Dr. Chaitan Khosla and at Stanford University. Under the guidance of my outstanding scientific advisory committee, I will obtain training essential to mastering enzymology, germ cell biology, and functional genomics. These research and career-development opportunities will empower me to establish an independent laboratory to study fundamental questions in the mammalian sperm-egg fertilization process at the levels of cell biology, mechanistic biochemistry, and structural biology.

项目概要/摘要 受精是一个重要的生物过程，涉及细胞表面识别、粘附、 单倍体精子和卵子融合形成新的、遗传上独特的二倍体生物体。出奇 人们对质膜如何相互作用和融合知之甚少，这主要是由于质膜的可用性有限 哺乳动物配子和捕获瞬时细胞外相互作用的技术挑战。 虽然配子膜融合机制仍然难以捉摸，但众所周知，人类 免疫球蛋白超家族蛋白 Izumo1 定位于精子的赤道段， 在配子融合发生之前，它与卵母细胞配体朱诺相互作用。迄今为止，出云1- Juno 是该通路中唯一确定的重要受体-配体对。最近的遗传学研究 小鼠已经发现了对男性生育能力至关重要的其他精子表面蛋白，但它们的相互作用 和监管尚未明确。 为了弥补这些主要差距，我围绕中心假设设计了一个研究计划 配子表面蛋白协调其结构重排和膜重塑， 导致配子融合。我将通过（1）确定分子机制来检验我的假设 人类 Izumo1 自组装，(2) 表征人类 Izumo 家族蛋白在 膜粘附和融合，以及（3）识别额外的受体-配体相互作用至关重要 用于配子融合和哺乳动物受精。我的多学科方法利用了高 解析结构分析以询问已知必需配子的功能组织 表面蛋白，并利用高通量正向遗传筛选来探索相互作用 其他新兴分子的网络。至关重要的是，理解这些相互作用将阐明 细胞与细胞识别和膜融合的机制，并揭示维持的基本因素 人类生育力和生殖健康。 从研究生院毕业，我接受了遗传学、细胞生物学和膜方面的广泛培训 生物化学。在我早期的博士后工作中，我磨练了细胞表面结构生物学的技能 病毒膜融合机的受体和蛋白质工程。我将发挥优秀 我的导师 Peter Kim 博士和我的合作导师 Chaitan 博士的实验室的研究环境 科斯拉和斯坦福大学。在我杰出的科学顾问的指导下 委员会，我将获得掌握酶学、生殖细胞生物学和功能学所必需的培训 基因组学。这些研究和职业发展机会将使我能够建立一个 研究哺乳动物精卵受精基本问题的独立实验室 细胞生物学、机械生物化学和结构生物学水平的过程。