An evolutionary and proteomic search for human gamete recognition proteins that mediate zona pellucida dissolution

介导透明带溶解的人类配子识别蛋白的进化和蛋白质组学研究

• 批准号: 10370974
• 负责人: WILLIE J SWANSON
• 金额: $ 22.93万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370974
• 关键词: 3-Dimensional; Address; Affinity; Binding; Biochemical; Bioinformatics; Biological Assay; Biology; Characteristics; Chemotaxis; Classification; Codon Nucleotides; Complementary DNA; Data; Databases; Detection; Distant; Egg Proteins; Event; Evolution; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; Female; Fertilization; Frequencies; Future; Genes; Genetic; Genomics; Germ Cells; Goals; Guanine + Cytosine Composition; Homologous Gene; Homologous Protein; Human; Human Genome; Knock-out; Length; Mass Spectrum Analysis; Measures; Mediating; Molecular; Morphology; Mus; N Domain; Oocytes; Performance; Protein Structure Initiative; Proteins; Proteome; Proteomics; RNA Splicing; Reproduction; Research; Resources; Specificity; Sperm Capacitation; Structure; Surface Plasmon Resonance; Technology; Testing; Testis; Transcript; Variant; Zona Pellucida; abalone; algorithm training; base; egg; env Gene Products; experience; extracellular; genetic approach; genomic data; improved; innovation; lysin; male; mammalian genome; molecular phenotype; nanopore; prediction algorithm; predictive test; reproductive; sperm cell; sperm protein; three dimensional structure; transcriptome; transcriptome sequencing; transcriptomics

Project Summary The identification of interacting male-female reproductive proteins is imperative for a molecular understanding of fertilization. The goal of this R21 proposal is to identify a set of human gamete recognition proteins using genomic and proteomic approaches, and then perform evolutionary guided computational bioinformatic searches for a homolog of abalone sperm lysin. It is known from three-dimensional protein crystallographic studies of the major proteins from the mammalian Zona Pellucida (ZP) and abalone Vitelline Envelope (VE) that they contain homologous ZP-N domains. Given the molecular homology of ZP-N domains and overall biochemical similarity of the egg coats, an obvious question to address is whether sperm use similar mechanisms (or homologous proteins) to generate a path through the egg coat. The dissolution of the abalone egg VE by the sperm protein lysin has been extensively characterized. However, it remains unknown how mammalian sperm penetrate egg coats, and no lysin homolog is currently identifiable in any mammalian genome. Our proposal address this question by (1) using long-read sequencing to generate deep coverage of full-length non-chimeric cDNAs for use in sperm proteomics, (2) using these resources as a database to perform detailed bioinformatic analyses aimed at generating a database of putative human gamete recognition proteins and identification of a human homolog of abalone sperm lysin and (3) functional tests of predicted protein's ability to disrupt the mouse ZP. If we identify a human sperm lysin, it will immediately open up new avenues of research into mammalian gamete recognition and how the human sperm passes the ZP. If we are unsuccessful at identifying a human lysin, we will have generated a valuable genomic and proteomic resource database for future studies of human sperm biology.

项目摘要 相互作用的男性生殖蛋白相互作用对于分子理解至关重要 受精。该R21提案的目的是使用 基因组和蛋白质组学方法，然后执行进化引导的计算生物信息学 寻找鲍鱼精子溶菌蛋白的同源物。它是从三维蛋白质晶体学知道的 研究来自哺乳动物Zona pellucida（ZP）和鲍鱼植物包膜（VE）的主要蛋白质的研究 它们包含同源ZP-N域。鉴于ZP-N结构域的分子同源性和总体 鸡蛋外套的生化相似性，一个明显的问题要解决的问题是精子是否使用类似 机制（或同源蛋白质）可以通过鸡蛋涂层生成一条路径。鲍鱼的解散 精子蛋白赖氨酸的鸡蛋VE已得到广泛的特征。但是，这仍然未知 哺乳动物的精子穿透卵外套，目前在任何哺乳动物中均无法识别溶菌蛋白 基因组。我们的建议通过（1）使用长阅读测序来解决这个问题 用于精子蛋白质组学的全长非核CDNA，（2）使用这些资源作为数据库 执行旨在生成假定人配子识别数据库的详细生物信息学分析 蛋白质和鲍鱼精子溶菌蛋白的人类同源物和（3）预测的功能测试 蛋白质破坏鼠标ZP的能力。如果我们识别人类精子溶菌蛋白，它将立即开放新的 哺乳动物配子识别的研究途径以及人类精子如何通过ZP。如果是 我们将产生宝贵的基因组和蛋白质组学资源，在识别人类赖氨酸方面失败了 数据库，用于未来的人类精子生物学研究。