Molecular cloning of a membrane receptor of maturation-inducing steroid hormone from fish oocytes
鱼卵母细胞成熟诱导类固醇激素膜受体的分子克隆
基本信息
- 批准号:09640798
- 负责人:
- 金额:$ 2.18万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:1997
- 资助国家:日本
- 起止时间:1997 至 1998
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Fish oocyte maturation is induced by maturation-inducing hormone, 17alpha, 20beta-dihydroxy-4-pregnen-3-one (l7alpha, 2Obeta-DP) acting upon an oocyte surface receptor.We evidenced that a pertussis toxin-sensitive inhibitory GTP-binding protein (Gi) was involved in the signal transduction pathway of l7alpha, 2Obeta-DP in rainbow trout oocytes.We cloned two Gialpha subunit cDNAs, Gialpha1 and Gialpha2, from Medaka (Oryzias latipes) oocyte cDNA library. Three rabbit polyclonal antibodies, A2, C4, and M1, were generated against deduced internal amino acid sequences of Gialpha1 and Gialpha2, and a common C-terminal sequence, respectively.Ml antibody successfully immunoprecipitated Gialphal and Gialpha2 subunit proteins.However, l7alpha, 2Obeta-DP-binding acitivity was almost not detected in the immunoprecipitate.l7alpha, 2Obeta-DP receptor activity was solubilized from membrane preparations of goldfish oocytes by sucrose monolaurate. Solubilized activity was serially separated by following four-step chromatography ; gel-filtration, anion-exchange, hydrophobic interaction, and lectin affinity chromatography.Major seven proteineous bands on SDS-PAGE were detected by silver staining.In medaka (Oryzias latipes) oocytes, the amount of l7alpha, 2Obeta-DP binding to the oocyte cortices was shown to increase with maturational competence of oocytes and reach a maximum one day before spawning.When oocyte maturation was triggered with l7alpha, 2Obeta-DP, 17alpha, 2Obeta-DP binding and Gi decreased quickly, suggesting a close relation between Gi and the l7alpha, 2Obeta-DP receptor.
通过成熟的激素,17Alpha,20beta-dihydroxy-4-4-Pregnen-3-One-3-One(L7Alpha,2obeta-dp)作用于卵母细胞受体对卵母细胞受体的作用。 L7Alpha,彩虹鳟鱼卵母细胞中的2obeta-dp。我们克隆了来自Medaka(Oryzias latipes)卵母细胞CDNA库的两个Gialpha亚基CDNA,Gialpha1和Gialpha2。分别针对定推的gialpha1和gialpha2的内部氨基酸序列以及一个常见的C末端序列而产生了三种兔多克隆抗体A2,C4和M1。在免疫沉淀。L7Alpha中,通过蔗糖单龙酸盐从金鱼卵母细胞的膜制剂中溶解2obeta-DP受体活性。溶解活性通过以下四步色谱分离。通过银色染色检测到SDS-PAGE上的七个蛋白质带,在Medaka(Oryzias latipes)卵母细胞中检测到凝胶效过,阴离子交换,疏水相互作用和凝集素亲和力色谱。产卵前一天最多。当卵母细胞成熟时,由L7Alpha,2obeta-dp,17alpha,17alpha,2obeta-DP结合和GI迅速降低,这表明GI与L7Alpha与L7Alpha之间的关系紧密相关。
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)
Yoshikuni M.: "Apertussis toxin sensitive GTP-binding protein is involved in the signal truns duction pathway of the maturation-inducinghormone(17α,20β-dihydroxy-4-pregnene-3-one)in fist oocytes." Steroids. (印刷中). (1999)
Yoshikuni M.:“百日咳毒素敏感的 GTP 结合蛋白参与拳头类固醇中成熟诱导激素 (17α,20β-二羟基-4-孕烯-3-一) 的信号传导途径。”按))(1999)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Yuichi Oba: "Inhibitory guanine-nucleotide-binding-regulatory protein α subunits in Medaka oocytes. cDNAcloning and decreased expression of proteinscturing oocyte maturation." European Journal of Biochemistry. 249. 846-853 (1997)
Yuichi Oba:“青鳉卵母细胞中的抑制性鸟嘌呤核苷酸结合调节蛋白 α 亚基。cDNA 克隆和卵母细胞成熟蛋白表达的降低。欧洲生物化学杂志 249。846-853 (1997)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Yoshikuni M.: "A PERTUSSIS TOXIN SENSITIVE GTP-BINDING PROTEIN IS INVOLVED IN THE SIGNAL TRANSDUCTION PATHWAY OF THE MATURATION-INDUCING HORMONE (17α,20β-DIHYDROXY-4-PREGNEN-3-ONE)IN FISH OOCYTES" Steroids. (印刷中). (1999)
Yoshikuni M.:“一种百日咳毒素敏感的 GTP 结合蛋白参与鱼卵母细胞中成熟诱导激素 (17α,20β-二羟基-4-PREGNEN-3-ONE) 的信号转导途径”(类固醇)。 1999)
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Yuichi Oba: "Inhibitory guanine-nucleotide-binding-regulortory protein α subunits in Medaka oocytes.cDNA cloning and decreased expression of proteins during oocyte maturation." European Journal of Biochemistry. 249. 846-853 (1997)
Yuichi Oba:“青鳉卵母细胞中的抑制性鸟嘌呤核苷酸结合调节蛋白 α 亚基。卵母细胞成熟过程中的 cDNA 克隆和蛋白质表达降低。” 249. 846-853 (1997)。
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- 影响因子:0
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YOSHIKUNI Michiyasu其他文献
YOSHIKUNI Michiyasu的其他文献
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{{ truncateString('YOSHIKUNI Michiyasu', 18)}}的其他基金
The mechanism of oocyte maturation in sea cucumber.
海参卵母细胞成熟机制.
- 批准号:
24570072 - 财政年份:2012
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
A study of neuronal differentiation of sea cucumber by the comprehensive analysis of neuropeptides.
神经肽综合分析研究海参神经元分化[J].
- 批准号:
20570057 - 财政年份:2008
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Study of maturation-inducing steroid hormone receptor of fish
鱼类成熟诱导类固醇激素受体的研究
- 批准号:
16570059 - 财政年份:2004
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Study of membrane steroid hormone receptor on fish oocyte maturation.
膜类固醇激素受体对鱼类卵母细胞成熟影响的研究。
- 批准号:
14540620 - 财政年份:2002
- 资助金额:
$ 2.18万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
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