Sequence-specific interaction between the disintegrin domain of mouse ADAM 3 and murine eggs: role of beta1 integrin-associated proteins CD9, CD81, and CD98.
Sequence-specific interaction between the disintegrin domain of mouse ADAM 3 and murine eggs: role of beta1 integrin-associated proteins CD9, CD81, and CD98.
复制标题
小鼠 ADAM 3 的解整合素结构域与小鼠卵之间的序列特异性相互作用：β1 整合素相关蛋白 CD9、CD81 和 CD98 的作用。
DOI：
10.1091/mbc.12.4.809
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发表时间：
2001
影响因子：
3.3
通讯作者：
White,JM
中科院分区：
文献类型：
--
作者：
Takahashi,Y;Bigler,D;Ito,Y;White,JM
ADAM 3是精子表面糖蛋白，与精卵粘附有关。由于对亚当斯的粘附活性知之甚少，我们研究了在细菌和昆虫细胞中制备的ADAM 3去整合素结构域与鼠卵的相互作用。这两种重组蛋白抑制精卵结合和融合的效力类似，我们最近报道的ADAM 2去整合素结构域。丙氨酸扫描诱变揭示了去整合素环7位谷氨酰胺的关键重要性。用结合到卵表面的ADAM 3去整合素结构域包被的荧光珠。珠结合被抑制的真实的，但不是由乱序，肽类似物的去整合素环。功能阻断性抗α6单克隆抗体（mAb）GoH 3也可抑制微珠结合，但非功能阻断性抗α6 mAb或抗αv或β3整联蛋白亚基的mAb均不能抑制微珠结合。我们还提供了证据表明，除了四跨膜蛋白CD 9外，其他两种β1-整合素相关蛋白，四跨膜蛋白CD 81以及单次跨膜蛋白CD 98在小鼠卵上表达。CD 9和CD 98抗体抑制体外受精和ADAM 3去整合素结构域的结合。我们的研究结果进行了讨论，在多个精子的亚当斯和多个卵子β1整合素相关蛋白的精卵结合和融合的参与。我们建议，鸡蛋表面的“tetraspan网”有利于受精，它可以这样做，通过促进ADAM整合素的相互作用。
ADAM 3 is a sperm surface glycoprotein that has been implicated in sperm-egg adhesion. Because little is known about the adhesive activity of ADAMs, we investigated the interaction of ADAM 3 disintegrin domains, made in bacteria and in insect cells, with murine eggs. Both recombinant proteins inhibited sperm-egg binding and fusion with potencies similar to that which we recently reported for the ADAM 2 disintegrin domain. Alanine scanning mutagenesis revealed a critical importance for the glutamine at position 7 of the disintegrin loop. Fluorescent beads coated with the ADAM 3 disintegrin domain bound to the egg surface. Bead binding was inhibited by an authentic, but not by a scrambled, peptide analog of the disintegrin loop. Bead binding was also inhibited by the function-blocking anti-α6 monoclonal antibody (mAb) GoH3, but not by a nonfunction blocking anti-α6 mAb, or by mAbs against either the αv or β3 integrin subunits. We also present evidence that in addition to the tetraspanin CD9, two other β1-integrin-associated proteins, the tetraspanin CD81 as well as the single pass transmembrane protein CD98 are expressed on murine eggs. Antibodies to CD9 and CD98 inhibited in vitro fertilization and binding of the ADAM 3 disintegrin domain. Our findings are discussed in terms of the involvement of multiple sperm ADAMs and multiple egg β1 integrin-associated proteins in sperm-egg binding and fusion. We propose that an egg surface “tetraspan web” facilitates fertilization and that it may do so by fostering ADAM–integrin interactions.