Arylsulfatase A is present on the pig sperm surface and is involved in sperm-zona pellucida binding
Arylsulfatase A is present on the pig sperm surface and is involved in sperm-zona pellucida binding
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DOI：
10.1006/dbio.2002.0690
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发表时间：
2002-07-01
影响因子：
2.7
通讯作者：
Tanphaichitr, N
中科院分区：
文献类型：
--
作者：
Carmona, E;Weerachatyanukul, W;Tanphaichitr, N
我们之前描述了猪精子表面蛋白 P68 与哺乳动物透明带 (ZP) 的亲和力。在本报告中，我们根据猪睾丸 AS-A cDNA 序列中 P68 胰蛋白酶肽序列的存在，将 P68 鉴定为芳基硫酸酯酶 A (AS-A)。我们的目标是证明精子表面存在 AS-A 并阐明其在 ZP 结合中的作用。免疫金电子显微镜显示精子表面存在 AS-A。此外，活猪精子和外周精子质膜蛋白提取物表现出AS-A的脱硫活性。值得注意的是，猪精子表面 AS-A 在 ZP 结合中的作用通过用抗 AS-A IgG/Fab 预处理精子后精子 ZP 结合的剂量依赖性减少以及 Alexa-430 缀合的精子表面 AS-A 与同源 ZP 的结合来证明。用抗猪ZP3抗体进行ZP预处理消除了AS-A结合，表明被识别为猪精子受体的ZP3是AS-A的结合配体。外源 ZP3 竞争性抑制 AS-A-ZP 结合的能力进一步证实了这一点。同样，纯化的 ZP3α（ZP3 的主要精子受体成分）对 AS-A-ZP 结合表现出巨大的抑制作用。所有这些结果都表明了 AS-A 在配子相互作用中的新功能。 (C) 2002 年爱思唯尔科学（美国）。
We have previously described the affinity of a pig sperm surface protein, P68, to mammalian zonae pellucidae (ZP). In this report, we identified P68 as arylsulfatase A (AS-A) based on the presence of P68 tryptic peptide sequences in the pig testis AS-A cDNA sequence. Our objective was to demonstrate the presence of AS-A on the sperm surface and to elucidate its role in ZP binding. Immunogold electron microscopy revealed the presence of AS-A on the sperm surface. Furthermore, live pig sperm and the extract of peripheral sperm plasma membrane proteins exhibited AS-A's desulfation activity. Significantly, the role of pig sperm surface AS-A in ZP binding was demonstrated by dose-dependent decreases of sperm-ZP binding upon sperm pretreatment with anti-AS-A IgG/Fab, and by the binding of Alexa-430-conjugated sperm surface AS-A to homologous ZP. ZP pretreatment with anti-pig-ZP3 antibody abolished AS-A binding, suggesting that ZP3, recognized as the pig sperm receptor, was AS-A's binding ligand. This was further confirmed by the ability of exogenous ZP3 to competitively inhibit AS-A-ZP binding. Similarly, purified ZP3alpha, a major sperm receptor component of ZP3, exhibited great inhibitory effect on AS-A-ZP binding. All of these results designated a new function of AS-A in gamete interaction. (C) 2002 Elsevier Science (USA).