SEQUENCE OF A RABBIT SPERM ZONA-PELLUCIDA BINDING-PROTEIN AND LOCALIZATION DURING THE ACROSOME REACTION
SEQUENCE OF A RABBIT SPERM ZONA-PELLUCIDA BINDING-PROTEIN AND LOCALIZATION DURING THE ACROSOME REACTION
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DOI：
10.1006/dbio.1994.1285
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发表时间：
1994-10-01
影响因子：
2.7
通讯作者：
ORAND, MG
中科院分区：
文献类型：
--
作者：
RICHARDSON, RT;YAMASAKI, N;ORAND, MG
哺乳动物精子与卵母细胞的细胞外基质或透明质酸的相互作用是导致成功受精的关键的第一步。在这种细胞-细胞外基质相互作用中，透明质酸的碳水化合物充当精子受体，精子表面提供凝集素样粘附分子。为了更好地了解精子与透明质酸的结合，我们分析了一种特异性的透明质酸结合蛋白（ZBP）。本研究确定了编码哺乳动物睾丸和精子特异性蛋白的mRNA序列，分子量为16，891 Da，我们将其命名为Sp17。对Sp17的分析表明，该mRNA存在于兔、小鼠和人的睾丸中，但不存在于任何测试的体细胞组织中。在兔中，Sp17是精子特异性自身抗原的兔精子自身抗原家族的17-kDa成员，由0.9和1.1 kb的两种mRNA编码。每个mRNA都有一个独特的5'非翻译区，但两者都有相同的编码区。推导的氨基酸序列的Sp17 ZBP显示了几个有趣的功能，包括人类睾丸cAMP依赖性蛋白激酶的N-末端的相似性。使用重组Sp17或Sp17肽G22 C的抗体在活精子上定位Sp17，发现Sp17的肽骨架在顶体反应开始之前是不可接近的。然而，在多聚甲醛固定，顶体完整的精子，肽骨架是接近的抗体定位Sp17的顶端表面。在家兔和其他类似物种中，放射冠（颗粒）细胞紧密粘附在透明卵丘上并合成β-糖蛋白，因此，放射冠精子可能已经在卵丘内开始了顶体反应。因此，兔精子表面将被修饰以在卵丘通过的最后阶段暴露Sp17多肽，因此Sp17将可用于初始的精子结合。本研究还表明，重组Sp17可以结合透明质酸，葡聚糖和硫酸葡聚糖。(C)1994年出版社出版。
The interaction of the mammalian spermatozoon with the oocyte's extracellular matrix or zona pellucida is a critical first step leading to successful fertilization. In this cell-extracellular matrix interaction it is the carbohydrate of the zona pellucida which serves as the sperm receptor and the surface of the spermatozoon which provides the lectin-like adhesion molecules. To better understand sperm-zona pellucida binding we have analyzed one specific zona binding protein (ZBP). This study has determined the mRNA sequence encoding a mammalian testis and sperm specific protein of 16,891 Da, which we have designated Sp17. Analysis of Sp17 revealed that the mRNA is present in rabbit, mouse, and human testes but not in any somatic tissue tested. In the rabbit, Sp17 is the 17-kDa member of the rabbit sperm autoantigen family of sperm specific autoantigens and is encoded by two mRNAs of 0.9 and 1.1 kb. Each mRNA has a unique 5' untranslated region but both have identical coding regions. The deduced amino acid sequence of the Sp17 ZBP showed several interesting features, including a similarity to the N-terminal of human testis cAMP-dependent protein kinase. Localization of Sp17 on live spermatozoa using antibodies to recombinant Sp17 or to the Sp17 peptide, G22C, revealed that the peptide backbone of Sp17 is inaccessible until the acrosome reaction begins. However, on paraformaldehyde fixed, acrosome intact spermatozoa, the peptide backbone is accessible to the antibodies which localize Sp17 to the apical surface. In the rabbit as well as other similar species in which the corona radiata (granulosa) cells adhere tightly to the zona pellucida and synthesize zona glycoproteins, the fertilizing spermatozoon may have already begun the acrosome reaction within the cumulus oophorus. Thus, the rabbit sperm surface would be modified to expose the Sp17 polypeptide during the final phase of cumulus passage and consequently Sp17 would be available for initial zona binding. The present study has also demonstrated that recombinant Sp17 can bind zona pellucida, dextran, and dextran sulfate. (C) 1994 Academic Press, Inc.