Receptor mediated yolk protein uptake in the crab Scylla serrata: crustacean vitellogenin receptor recognizes related mammalian serum lipoproteins
Receptor mediated yolk protein uptake in the crab Scylla serrata: crustacean vitellogenin receptor recognizes related mammalian serum lipoproteins
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DOI：
10.1002/mrd.10106
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发表时间：
2002-04
影响因子：
2.5
通讯作者：
S. Warrier;T. Subramoniam
中科院分区：
文献类型：
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作者：
S. Warrier;T. Subramoniam
受体介导的主要卵黄蛋白前体卵黄蛋白原（Vg）的摄取对产蛋动物的卵母细胞生长至关重要。本研究从锯缘青蟹（Scylla serrata）卵母细胞中分离出Vg质膜受体。使用从大鼠分离的标记的蟹Vg（125 I-Vg）以及标记的低密度脂蛋白（125 I-LDL）和极低密度脂蛋白（125 I-VLDL），通过配体印迹法可视化卵黄原蛋白受体（VgR）蛋白。通过免疫电镜观察，发现Vg在卵母细胞中具有内吞作用。通过凝胶过滤高效液相色谱（HPLC）纯化Vg受体，其分子量估计为230 kDa。在直接结合研究中，该受体对螃蟹Vg表现出高亲和力（解离常数Kd 0.8 × 10−6 M）。在Ca ~（2+）存在下，卵黄蛋白原受体与蟹Vg的亲和力增加，而苏拉明可抑制这种结合，表明蟹Vg受体与低密度脂蛋白受体（LDLR）超家族的受体蛋白具有相似性。此外，螃蟹VgR显示出显着的结合能力，哺乳动物致动脉粥样硬化脂蛋白，如LDL和VLDL。这表明无脊椎动物（蟹）Vg和脊椎动物（大鼠）LDL和VLDL之间的受体结合位点是紧密保守的。摩尔Reprod. Dev. 61：536-548，2002.© 2002 Wiley利斯公司
The receptor‐mediated uptake of major yolk protein precursor, vitellogenin (Vg) is crucial for oocyte growth in egg laying animals. In the present study plasma membrane receptor for Vg was isolated from the oocyte of the red mud crab, Scylla serrata. Vitellogenin receptor (VgR) protein was visualized by ligand blotting using labeled crab Vg (125I‐Vg) as well as labeled low density lipoprotein (125I ‐LDL) and very low density lipoprotein (125I‐VLDL) isolated from rat. The endocytosis of Vg was visualized in the crab oocyte by ultrastructural immunolocalization of Vg. The Vg receptor was purified by gel filtration high performance liquid chromatography (HPLC) and its molecular weight was estimated to be 230 kDa. In direct binding studies, the receptor exhibited high affinity (dissociation constant Kd 0.8 × 10−6 M) for crab Vg. Vitellogenin receptor was observed to have an increased affinity to crab Vg in the presence of Ca2+ and the binding was inhibited by suramin, suggesting similarities between crab VgR and low density lipoprotein receptor (LDLR) superfamily of receptor protein. Furthermore, the crab VgR showed significant binding ability to mammalian atherogenic lipoproteins such as LDL and VLDL. This suggests that there is a tight conservation of receptor binding sites between invertebrate (crab) Vg and vertebrate (rat) LDL and VLDL. Mol. Reprod. Dev. 61:536–548, 2002. © 2002 Wiley‐Liss, Inc.