Isolation of protein from Urechis sperm acrosomal granules that binds sperm to eggs and initiates development
Isolation of protein from Urechis sperm acrosomal granules that binds sperm to eggs and initiates development
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从 Urechis 精子顶体颗粒中分离蛋白质，该颗粒将精子与卵子结合并启动发育
DOI：
10.1016/0012-1606(86)90373-8
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发表时间：
1986
影响因子：
2.7
通讯作者：
L. Holland
中科院分区：
文献类型：
--
作者：
M. Gould;J. Stephano;L. Holland
蛋白质是从卵子表面紧密结合的环状顶体颗粒中分离出来的，也启动了发育。顶体蛋白在醋酸-尿素PAGE中以单一主带迁移，赖氨酸+精氨酸含量为∼的50%，缺少碳水化合物。十六烷基三甲基溴化铵PAGE显示顶体蛋白的相对分子质量为25,000-30,000 Da。用荧光胺选择性地标记顶体反应精子的顶体环，荧光在整个分离过程中持续存在，并在凝胶上的主带中观察到。尽管顶体蛋白和尿素精蛋白具有相似的氨基酸含量和在醋酸-尿素凝胶中的迁移，但顶体蛋白与鱼精蛋白的不同之处在于，它在热的5%三氯乙酸和冷的0.25NH_2SO_4中相对不溶，它在十六烷基溴化铵和四甲基溴化铵凝胶中的迁移，以及在热裂解酶消化后在其肽图上的一个主要位置。尾丝虫顶体蛋白对虫卵的凝集不具有种特异性，包括各种棘皮动物的虫卵和藻类受精卵以及尾丝虫卵。顶体蛋白制剂和从凝胶主带中提取的蛋白质都启动了尾端卵子的发育，导致卵子变圆、表面被隆起、生发泡破裂、极体形成和多精块的建立。多聚赖氨酸和小牛胸腺组蛋白在激活海胆卵子方面同样有效，但尿裂精子鱼精蛋白的效果较差，而鲑鱼精子鱼精蛋白虽然高度碱性，但只激活一小部分卵子。尿裂精子顶体蛋白还诱导海胆卵子受精膜的部分升高，类似于海胆卵子对尿裂精子的反应。海胆卵不被多聚赖氨酸激活。
Protein was isolated from the ring-shaped acrosomal granules ofUrechissperm which bound tightly toUrechisegg surface coats and also initiated development. The acrosomal protein preparation migrated as a single major band in acetic acid-urea PAGE, had a lysine + arginine content of ∼50%, and lacked carbohydrate. The molecular mass of the acrosomal protein was 25,000–30,000 Da in cetyltrimethylammonium bromide PAGE. Acrosomal rings of acrosomereacted sperm were selectively labeled with fluorescamine and the fluorescence persisted throughout the isolation procedure and was observed in the major band on gels. Although the acrosomal protein andUrechissperm protamine had similar amino acid contents and migrated similarly in acetic acid-urea gels, acrosomal protein differed from protamine by its relative insolubility in hot 5% trichloroacetic acid and cold 0.25NH2SO4, by its migration in cetyl- and tetratrimethylammonium bromide PAGE and in a major spot on its peptide map following thermolysin digestion. Agglutination of eggs byUrechisacrosomal protein was not species-specific and included various echinoderm eggs and algal zygotes as well asUrechiseggs. Both the acrosomal protein preparation and protein extracted from the major band of gels initiated development ofUrechiseggs, causing rounding out, elevation of the surface coat, germinal vesicle breakdown, polar body formation, and establishment of a polyspermy block. Polylysine and calf thymus histones were equally effective in activatingUrechiseggs, butUrechissperm protamine was less effective, and salmon sperm protamine, although highly basic, activated only a small percentage of eggs.Urechisacrosomal protein also induced partial elevation of sea urchin egg fertilization envelopes, similar to the response of sea urchin eggs toUrechissperm. Sea urchin eggs were not activated by polylysine.