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首页> 外文期刊>Journal of cellular physiology. >Study on the role of calmodulin in sperm function through the enrichment and identification of calmodulin-binding proteins in bovine ejaculated spermatozoa
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Study on the role of calmodulin in sperm function through the enrichment and identification of calmodulin-binding proteins in bovine ejaculated spermatozoa

机译：研究钙调蛋白在精子功能的作用通过浓缩和识别在牛射精calmodulin-binding蛋白质精子

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Calmodulin is a small, highly conserved acidic protein present at high levels in spermatozoa that mediates numerous intracellular Ca~(2+)-dependent events. Sperm motility and fertilizing ability results from an array of biochemical pathways under Ca~(2+) control, in which the importance of calmodulin is not fully understood. The role of calmodulin in sperm function has been mostly assessed using antagonists. Nevertheless, few known calmodulin-regulated enzymes have been described in spermatozoa regarding their involvement in sperm function. To further understand the role of this important Ca~(2+) mediator in spermatozoa, different studies were also undertaken to investigate and to identify sperm calmodulin-binding proteins and determine their localization and subcellular distribution as an attempt to elucidate the role of this important Ca~(2+) mediator. In the present study, sperm calmodulin-binding proteins were identified by mass spectrometry after Ca~(2+)-dependent biotinylated-calmodulin binding on sperm head proteins subjected to 2D electro-phoresis and transferred on a polyvinylidene difluoride membrane. Calmodulin binding protein identification was also done on detergent extracted whole sperm proteins pulled down in a Ca~(2+)-dependent manner by calmodulin-conjugated sepharose beads. In this latter group, 300 proteins were identified in at least two experiments out of three, and those identified in the three independent experiments were analyzed for over re presented biological processes using the Bos taurus Gene Ontology database. Proteins with known function in reproductive processes, fertilization, sperm-egg recognition, sperm binding to the zona pellucida, regulation of sperm capacitation, and sperm motility were identified and further emphasize the importance of calmodulin in sperm function.
机译：钙调蛋白是一种小,高度保守的酸性蛋白在精子的高水平产生大量的细胞内Ca ~(2 +)端依赖事件。施肥结果数组的能力生化途径在Ca ~(2 +)控制钙调蛋白的重要性是不完全呢理解。功能主要是评估使用拮抗剂。calmodulin-regulated酶已被描述在精子有关他们的参与精子的功能。这个重要的Ca ~(2 +)中介在精子,不同的研究也进行调查和识别精子calmodulin-binding蛋白质和决定他们的本地化和亚细胞分布试图阐明这一重要的角色Ca ~(2 +)中介。calmodulin-binding蛋白质被确定质谱在Ca ~(2 +)端依赖biotinylated-calmodulin绑定在精子的头上二维电泳和蛋白质转移在聚乙二烯二氟化物膜。识别也在洗涤剂提取全部推倒在精子蛋白质Ca ~(2 +)由calmodulin-conjugated端依赖的方式琼脂糖珠。蛋白质在至少两个标识实验三,和那些中确定三个独立的实验进行了分析了通过生物过程呈现出来牛基因本体数据库。与已知函数在生殖过程中,施肥、精卵识别、精子绑定到透明带,调节精子获能的精子,精子的运动性识别并进一步强调的重要性钙调蛋白在精子的功能。

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