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Structure and function of epididymal protein cysteine-rich secretory protein-1 被引量:4
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作者 Kenneth P. Roberts Daniel S. Johnston +5 位作者 Michael A. Nolan Joseph L. Wooters Nicole C. Waxmonsky Laura B. Piehl Kathy M. Ensrud-Bowlin David W. Hamilton 《Asian Journal of Andrology》 SCIE CAS CSCD 2007年第4期508-514,共7页
Cysteine-rich secretory protein-1 (CRISP-1) is a glycoprotein secreted by the epididymal epithelium. It is a member of a large family of proteins characterized by two conserved domains and a set of 16 conserved cyst... Cysteine-rich secretory protein-1 (CRISP-1) is a glycoprotein secreted by the epididymal epithelium. It is a member of a large family of proteins characterized by two conserved domains and a set of 16 conserved cysteine residues. In mammals, CRISP-1 inhibits sperm-egg fusion and can suppress sperm capacitation. The molecular mechanism of action of the mammalian CRISP proteins remains unknown, but certain non-mammalian CRISP proteins can block ion channels. In the rat, CRISP-1 comprises two forms referred to as Proteins D and E. Recent work in our laboratory demonstrates that the D form of CRISP-1 associates transiently with the sperm surface, whereas the E form binds tightly. When the spermatozoa are washed, the E form of CRISP-1 persists on the sperm surface after all D form has dissociated. Cross-linking studies demonstrate different protein-protein interaction patterns for D and E, although no binding partners for either protein have yet been identified. Mass spectrometric analyses revealed a potential post-translational modification on the E form that is not present on the D form. This is the only discernable difference between Proteins D and E, and presumably is responsible for the difference in behavior of these two forms of rat CRISP- 1. These studies demonstrate that the more abundant D form interacts with spermatozoa transiently, possibly with a specific receptor on the sperm surface, consistent with a capacitation-suppressing function during sperm transit and storage in the epididymis, and also confirm a tightly bound population of the E form that could act in the female reproductive tract. (Asian J Androl 2007 July; 9: 508-514) 展开更多
关键词 cysteine-rich secretory protein-1 EPIDIDYMIS SPERM CAPACITATION
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小鼠富含半胱氨酸分泌蛋白-1真核表达载体的构建及其在COS-7细胞中的表达 被引量:4
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作者 罗金 杨菁 +1 位作者 徐望明 尹太郎 《生殖与避孕》 CAS CSCD 北大核心 2010年第8期509-513,共5页
目的:构建小鼠富含半胱氨酸分泌蛋白-1(Crisp-1)真核表达载体,并观察其在COS-7细胞中的表达。方法:从小鼠附睾组织中提取总RNA,RT-PCR扩增Crisp-1基因开放阅读框(ORF),将目的基因克隆至真核表达载体pcDNA3.1上,经脂质体介导转染COS-7细... 目的:构建小鼠富含半胱氨酸分泌蛋白-1(Crisp-1)真核表达载体,并观察其在COS-7细胞中的表达。方法:从小鼠附睾组织中提取总RNA,RT-PCR扩增Crisp-1基因开放阅读框(ORF),将目的基因克隆至真核表达载体pcDNA3.1上,经脂质体介导转染COS-7细胞,并利用间接免疫荧光和RT-PCR检测其在COS-7细胞中的表达。结果:酶切电泳鉴定结果显示Crisp-1基因克隆入载体中,测序结果证实克隆的基因序列与基因库(GeneBank)中的Crisp-1序列相符。脂质体转染后,间接免疫荧光和RT-PCR结果均表明重组质粒pcDNA3.1-Crisp-1能在COS-7细胞中表达。结论:成功获得了能在COS-7细胞中表达的小鼠Crisp-1DNA疫苗的真核表达质粒pcDNA3.1-Crisp-1,为后续研究其生物学活性及免疫避孕效应奠定了基础。 展开更多
关键词 半胱氨酸分泌蛋白-1(crisp-1) 真核表达载体 COS-7细胞 避孕疫苗
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小鼠富含半胱氨酸分泌蛋白-1DNA疫苗免疫避孕效果的评价 被引量:4
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作者 罗金 杨菁 +3 位作者 程琰 李维 李星 徐望明 《生殖与避孕》 CAS CSCD 2013年第3期145-148,172,共5页
目的:观察小鼠富含半胱氨酸分泌蛋白-1(Crisp-1)DNA避孕疫苗免疫雌、雄性BALB/c小鼠后特异性抗体的表达及免疫避孕效果。方法:将雌、雄性BALB/c小鼠随机分为实验组和对照组,每组雌鼠12只,雄鼠8只。每只动物经肌肉接种重组质粒pcDNA3.1-C... 目的:观察小鼠富含半胱氨酸分泌蛋白-1(Crisp-1)DNA避孕疫苗免疫雌、雄性BALB/c小鼠后特异性抗体的表达及免疫避孕效果。方法:将雌、雄性BALB/c小鼠随机分为实验组和对照组,每组雌鼠12只,雄鼠8只。每只动物经肌肉接种重组质粒pcDNA3.1-Crisp-1或pcDNA3.1空载体3次,ELISA测定小鼠血清中抗Crisp-1抗体水平的变化,Western blotting检测抗体的特异性。免疫结束后2周,将实验小鼠分别与未接种的正常雌、雄性小鼠合笼,记录每组雌鼠的妊娠率与每窝产仔数。结果:重组质粒pcDNA 3.1-Crisp-1可以在小鼠体内诱发特异性抗Crisp-1免疫应答,免疫后雌、雄性小鼠妊娠率和平均每窝产仔数均显著降低(P<0.05)。结论:重组质粒pcDNA3.1-Crisp-1具有一定的抗生育潜能。 展开更多
关键词 富含半胱氨酸分泌蛋白-1(crisp-1) DNA疫苗 免疫效果
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