CHARACTERIZATION OF SPERM MEMBRANE AUTOANTIGENS

精子膜自身抗原的表征

• 批准号: 3312527
• 负责人: MICHAEL GENE ORAND
• 金额: $ 13.15万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-01 至 1997-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3312527
• 关键词: acrosin; acrosome; arylsulfatases; autoantibody; autoantigens; binding proteins; cell membrane; chemical binding; complementary DNA; digital imaging; egg /ovum; electron microscopy; fertility immunology; fertilization; genetic manipulation; laboratory mouse; laboratory rabbit; messenger RNA; molecular cloning; molecular site; monoclonal antibody; nucleoproteins; protein purification; protein sequence; protein structure function; radiotracer; sperm; surface antigens

The long term goals of this research project are to define both chemically and biologically a set of sperm molecules which are necessary for one or more steps in the fertilization process. During the current grant period emphasis has been placed on defining the biological function of the rabbit sperm membrane autoantigen, RSA, as a zona pellucida binding protein and on obtaining CDNA clones which would allow further characterization of sperm autoantigenic sites and zona binding proteins. To continue this line of investigation, this revised renewal application will focus on four sperm molecules selected for study at the molecular level with regard to their role in fertilization and zona binding. Using the cyclic model as a working hypothesis, which was developed during the current grant period and which is explained in detail in Section C.4.(b).1-4, this proposal will specifically investigate the mechanism by which spermatoza attach to and penetrate (degrade) the zona pellucida. An understanding of the mechanism of sperm-zona interaction would allow precise targets for both infertility diagnosis and contraception. Therefore, this proposal's specific aims are: I. The mechanism of action of three sperm molecules on binding to and interacting with (degrading or releasing) the zona pellucida. I.1) The molecular characterization of the rabbit sperm membrane autoantigen, RSA, with regard to the amino acid sequences which specifically bind zona pellucida. I.2) The molecular characterization of the role of the sperm enzyme acrosin in zona binding. 3) The molecular characterization of the role of the sperm enzyme arylsulfatase A in zona binding. II. The isolation and biological characterization of a newly discovered sperm nuclear autoantigenic protein (NASP), with regard to: (A) biological function and (B) the amino acid sequences of autoantibody binding sites. Molecular sites methodology will be used to characterize specific sequences.

该研究项目的长期目标是定义 从化学和生物学上一组必要的精子分子 对于受精过程中的一个或多个步骤。在电流期间 赠款时期的重点是定义生物学 兔子精子膜自动抗原的功能，RSA，作为zona 蛋白质结合蛋白和获得cDNA克隆，这将允许 精子自身抗原位点和Zona结合的进一步表征 蛋白质。为了继续进行这一调查，此修订了续约 应用将集中于选择用于研究的四个精子分子 分子水平关于它们在受精和Zona中的作用 结合。使用环状模型作为工作假设，这是 在当前赠款期间开发，这在 C.4。（b）.1-4节的详细信息，该提案将专门 研究精子附着并穿透的机制 （退化）Zona Pellucida。对机制的理解 精子 - 佐纳的互动将允许两种不育的精确目标 诊断和避孕。因此，该提议的具体目的 是：I。三个精子分子在结合到与的作用机理 并与（降解或释放）Zona Pellucida相互作用。 I.1） 兔精子自动抗原的分子表征， RSA，关于特异性结合的氨基酸序列 Zona Pellucida。 i.2）分子表征的作用 Zona结合中的精子酶Acrosin。 3）分子表征 精子酶芳基硫酸酶A在Zona结合中的作用。 ii。这 新发现的精子的隔离和生物学表征 关于：（a）生物学的核自身抗原蛋白（NASP） 功能和（b）自身抗体结合位点的氨基酸序列。 分子位点方法将用于表征特定 序列。