PURIFICATION AND MOLECULAR-PROPERTIES OF VITELLIN FROM THE SILKWORM, BOMBYX-MORI
PURIFICATION AND MOLECULAR-PROPERTIES OF VITELLIN FROM THE SILKWORM, BOMBYX-MORI
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DOI：
10.1016/0020-1790(80)90074-8
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发表时间：
1980-01-01
期刊：
影响因子：
--
通讯作者：
CHINO, H
中科院分区：
文献类型：
--
作者：
IZUMI, S;TOMINO, S;CHINO, H
用一种简单的方法从家蚕卵中提纯卵黄蛋白，该方法包括低离子浓度下pH=6的特异性沉淀和DEAE-纤维素柱层析。经凝胶电泳法、EM法和超速离心法测定，最终制备的产物均一性较好。卵黄蛋白被定义为糖脂蛋白，其沉降系数(S20，W)为13.5s，相对分子质量为44万。分子呈近球形，直径约13 nm。该分子含有3%的甘露糖和7.5%的总脂，包括三酰甘油、二甘油、胆固醇、磷脂酰胆碱和磷脂酰乙醇胺。氨基酸组成中谷氨酸和天冬氨酸含量较高，蛋氨酸含量较低。该分子由两个不同的亚基组成，其相对分子质量分别为180,000和42,000，天然分子为四聚体，每个亚基由2个分子组成。实现了两个亚基的分离，甘露糖只与较重的亚基共价结合。兔抗卵黄蛋白抗体与血淋巴卵黄原蛋白发生交叉反应，但不与其他血淋巴蛋白发生交叉反应，也不与蝗虫卵黄原蛋白发生交叉反应。该抗体还与家蚕血淋巴卵黄原蛋白发生反应。
Vitellin was purified from the eggs of the silkworm B. mori by a simple method which included a specific precipitation at pH 6 under low ionic concentration and DEAE-cellulose column chromatography. The final preparation was highly homogeneous as judged by gel electrophoresis, EM and ultracentrifugation. Vitellin was defined as glycolipoprotein with a sedimentation coefficient (S20,W) of 13.5S and a MW of 440,000. The molecule was almost spherical in shape with a diameter of 13 nm. The molecule contained 3% mannose and 7.5% total lipids which comprised triacylglycerol, diacylglycerol, cholesterol, phosphatidylcholine and phosphatidylethanolamine. The amino acid composition displayed a high content of glutamic and aspartic acids and a low content of methionine. The molecule was composed of 2 non-identical subunits with MW of 180,000 and 42,000, and the native molecule was assumed to be a tetramer composed of 2 molecules of each of these subunits. Separation of the 2 subunits was achieved, and mannose was covalently associated only with the heavier subunit. The rabbit anti-egg vitellin antibody cross-reacted with the hemolymph vitellogenin but not with other hemolymph proteins, nor with the vitellogenin from Locusta migratoria. The antibody also reacted with the hemolymph vitellogenin of the silkworm Philosamia cynthia.