Identification of guanylate cyclases and related signaling proteins in sperm tail from sea stars by mass spectrometry.
Identification of guanylate cyclases and related signaling proteins in sperm tail from sea stars by mass spectrometry.
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通过质谱法鉴定海星精子尾部中的鸟苷酸环化酶和相关信号蛋白。
DOI：
10.1007/s10126-008-9096-7
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Moriyama,Hideaki
中科院分区：
文献类型：
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作者：
Nakachi,Mia;Matsumoto,Midori;Terry,PhilipM;Cerny,RonaldL;Moriyama,Hideaki
海洋无脊椎动物利用体外受精来利用有性生殖的优势，是一种极好的生存策略。为了防止错配，只有在受精过程中经过严格规定的步骤后才能成功受精。在海星中，受精过程从精子的趋化开始，随后精子到达卵子外壳时过度激活，然后精子穿透卵子外壳，实现融合。为了探讨趋化性及以下信号传导的启动是否具有物种特异性，我们在海星、Asterias amurensis、A。紫杉，和木犀草。由于信使核糖核酸（mRNA）的转录在配子中受到抑制，因此不能通过检测mRNA谱来研究精子蛋白在受精过程中的作用。因此，本研究采用质谱法进行蛋白质组学分析。在海星中，在接受类固醇精子激活肽（asterosap）后，精子尾部的受体膜结合鸟苷酸环化酶触发精子趋化。我们证实了三种海星中存在膜结合的鸟苷酸环化酶，它们都具有相同的结构域，包括胞外结构域、激酶样结构域和鸟苷酸环化酶结构域。大部分肽来自于分布在蛋白质溶剂侧的α-螺旋。更多的肽从细胞内区域被回收。跨膜结构域还没有恢复。受体的功能似乎在物种之间是保守的。此外，我们确定了可能参与鸟苷酸环化酶触发的信号通路的蛋白质。
Marine invertebrates employ external fertilization to take the advantages of sexual reproduction as one of excellent survival strategies. To prevent mismatching, successful fertilization can be made only after going though strictly defined steps in the fertilization. In sea stars, the fertilization process starts with the chemotaxis of sperm followed by hyperactivation of sperm upon arriving onto the egg coat, and then sperm penetrate to the egg coat before achieving the fusion. To investigate whether the initiation of chemotaxis and the following signaling has species specificity, we conducted comparative studies in the protein level among sea stars,Asterias amurensis,A. forbesi, andAsterina pectinifera. Since transcription of messenger ribonucleic acid (mRNA) has been suppressed in gamete, the roles of sperm proteins during the fertilization cannot be investigated by examining the mRNA profile. Therefore, proteomics analysis by mass spectrometry was used in this study. In sea stars, upon receiving asteroidal sperm-activating peptide (asterosap), the receptor membrane-bound guanylate cyclases in the sperm tail trigger sperm chemotaxis. We confirmed the presence of membrane-bound guanylate cyclases in the three sea star species, and they all had the same structural domains including the extracellular domain, kinase-like domain, and guanylate cyclase domain. The majority of peptides recovered were from α-helices distributed on the solvent side of the protein. More peptides were recovered from the intracellular domains. The transmembrane domain has not been recovered. The functions of the receptors seemed to be conserved among the species. Furthermore, we identified proteins that may be involved in the guanylate cyclase-triggered signaling pathway.