Glycosyltransferase function during fertilization
受精过程中糖基转移酶的功能
基本信息
- 批准号:6708068
- 负责人:
- 金额:$ 34.2万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1996
- 资助国家:美国
- 起止时间:1996-12-01 至 2007-02-28
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
DESCRIPTION (provided by applicant): My laboratory is interested in the molecular basis of cellular interactions during mammalian fertilization. During the past 20 years, we have used a variety of biochemical and molecular approaches to characterize an unusual sperm surface receptor (Beta-1,4-galactosyltransferase I, GaIT) for glycoside ligands in the egg zona pellucida. With the exception of some early work, HD 23479 has supported all of our studies of GaIT function during fertilization.
GaIT is traditionally viewed as a biosynthetic enzyme in the Golgi complex, although a small amount also appears on the surface of somatic cells. Sperm are unusual in that all of their GaIT is expressed on the sperm surface, where it selectively binds to the egg coat glycoprotein, ZP3. In response to ZP3-induced aggregation, GaIT activates, directly or indirectly, a pertussis toxin-sensitive G-protein cascade, contributing to the acrosome reaction. Our recent studies have examined the consequences of altering GaIT expression, both positively and negatively, on heterologous cells and on mouse sperm. In this regard, Xenopus oocytes expressing murine GaIT bind ZP3, and aggregation of GaIT by ZP3 or anti-GaIT antibodies induces G-protein activation and subsequent activation of the Xenopus oocyte. Similarly, sperm that overexpress GaIT bind more ZP3 and undergo accelerated G-protein activation and acrosome reactions. In contrast, sperm that are null for GaIT do not bind ZP3 and do not undergo ZP3-dependent acrosome reactions. These results show clearly that Gall functions as a ZP3 receptor and is capable of activating a pertussis toxin-sensitive G-protein cascade.
Despite the fact that GaIT-null sperm do not bind ZP3, they still bind to the zona pellucida, although they do not undergo subsequent zona-induced acrosomal exocytosis. This implies that non-GalT:non-ZP3 receptor:ligand interactions mediate initial sperm-zona adhesion, and that GalT-ZP3 interactions participate in subsequent acrosomal exocytosis. The specific Aims of this proposal will examine candidates for these newly appreciated sperm receptors and egg coat ligands. In the first Aim, we will test the hypothesis that the egg coat ligand responsible for sperm adhesion is a ZP3-independent, peripherally associated component of the egg coat. The second Aim will test whether the sperm protein p47 functions as a binding protein for the egg coat. Finally, in the third Aim, we will continue our analysis of GalT-mediated signal transduction leading to the acrosome reaction.
描述(由申请人提供):我的实验室对哺乳动物受精过程中细胞相互作用的分子基础感兴趣。在过去的20年中,我们已经使用了各种生化和分子方法来表征一个不寻常的精子表面受体(β-1,4-半乳糖基转移酶I,GaIT)的糖苷配体在卵透明质酸。除了一些早期的工作,HD 23479支持了我们所有关于受精过程中GaIT功能的研究。
GaIT传统上被认为是高尔基复合体中的生物合成酶,尽管少量也出现在体细胞表面。精子是不寻常的,因为它们所有的GaIT都表达在精子表面,在那里它选择性地与卵膜糖蛋白ZP 3结合。响应于ZP 3诱导的聚集,GaIT直接或间接地激活百日咳毒素敏感的G蛋白级联反应,促进顶体反应。我们最近的研究已经检查了改变GaIT表达的后果,无论是积极的还是消极的,对异源细胞和小鼠精子。在这方面,表达鼠GaIT的非洲爪蟾卵母细胞结合ZP 3,并且GaIT通过ZP 3或抗GaIT抗体的聚集诱导G蛋白活化和随后的非洲爪蟾卵母细胞的活化。同样,过表达GaIT的精子结合更多的ZP 3,并经历加速的G蛋白活化和顶体反应。相反,GaIT无效的精子不结合ZP 3,也不发生ZP 3依赖性顶体反应。这些结果清楚地表明,Gall作为ZP 3受体发挥功能,并能够激活百日咳毒素敏感的G蛋白级联反应。
尽管事实上,步态无效的精子不结合ZP 3,他们仍然结合透明带,虽然他们不经历随后的透明带诱导的顶体胞吐。这意味着非GalT:非ZP 3受体:配体相互作用介导初始精子粘附,并且GalT-ZP 3相互作用参与随后的顶体胞吐。本提案的具体目的是研究这些新发现的精子受体和卵膜配体的候选者。在第一个目标,我们将测试的假设,负责精子粘附的卵外套配体是一个ZP 3独立的,外围相关的组件的卵外套。第二个目的是测试精子蛋白p47是否作为卵被的结合蛋白。最后,在第三个目标中,我们将继续分析半乳糖苷酶介导的信号转导导致顶体反应。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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BARRY D SHUR其他文献
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{{ truncateString('BARRY D SHUR', 18)}}的其他基金
GLYCOSYLTRANSFERASE FUNCTION DURING FERTILIZATION
受精过程中的糖基转移酶功能
- 批准号:
2888952 - 财政年份:1996
- 资助金额:
$ 34.2万 - 项目类别:
Glycosyltransferase function during fertilization
受精过程中糖基转移酶的功能
- 批准号:
6864810 - 财政年份:1996
- 资助金额:
$ 34.2万 - 项目类别:
Glycosyltransferase function during fertilization
受精过程中糖基转移酶的功能
- 批准号:
6542488 - 财政年份:1996
- 资助金额:
$ 34.2万 - 项目类别:
Glycosyltransferase function during fertilization
受精过程中糖基转移酶的功能
- 批准号:
7028283 - 财政年份:1996
- 资助金额:
$ 34.2万 - 项目类别:
GLYCOSYLTRANSFERASE FUNCTION DURING FERTILIZATION
受精过程中的糖基转移酶功能
- 批准号:
2408085 - 财政年份:1996
- 资助金额:
$ 34.2万 - 项目类别:
Glycosyltransferase function during fertilization
受精过程中糖基转移酶的功能
- 批准号:
6640123 - 财政年份:1996
- 资助金额:
$ 34.2万 - 项目类别:
GLYCOSYLTRANSFERASE FUNCTION DURING FERTILIZATION
受精过程中的糖基转移酶功能
- 批准号:
6182226 - 财政年份:1996
- 资助金额:
$ 34.2万 - 项目类别:
GLYCOSYLTRANSFERASE FUNCTION DURING FERTILIZATION
受精过程中的糖基转移酶功能
- 批准号:
2673556 - 财政年份:1996
- 资助金额:
$ 34.2万 - 项目类别:
Training Pgm in Biochemistry, Cell and Molecular Biology
生物化学、细胞和分子生物学培训 Pgm
- 批准号:
6845065 - 财政年份:1990
- 资助金额:
$ 34.2万 - 项目类别:
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