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Fibrinogen‐related protein, FGL2, of hamster cauda epididymal fluid: Purification, kinetic analysis of its prothrombinase activity, and its role in segregation of nonviable spermatozoa
Molecular Reproduction and Development ( IF 2.7 ) Pub Date : 2020-11-20 , DOI: 10.1002/mrd.23438
Subir K Nagdas 1 , Shamar Wallace 1 , Don Eaford 1 , Rashad Baker 1 , Ky'ara Carr 1 , Samir S Raychoudhuri 2
Affiliation  

Although the epididymal environment promotes the maturation and survival of spermatozoa, not all spermatozoa remain viable during passage through the epididymis. Does the epididymis has a protective mechanism(s) to segregate the viable sperm from defective spermatozoa? Previously, we identified 260/280 kDa oligomers (termed eFGL—Epididymal Fibrinogen‐Like oligomer) are composed of two disulfide‐linked subunits: a 64 kDa polypeptide identified as fibrinogen‐like protein‐2 (FGL2) and a 33 kDa polypeptide identified as fibrinogen‐like protein‐1 (FGL1). Our morphological studies demonstrated that the eFGL, secreted from the principal cells of the cauda epididymis, is polymerized into a death cocoon‐like complex (DCF), masking defective luminal spermatozoa but, not the viable sperm population. In the present study, we purified FGL2 from hamster cauda epididymal fluid toward homogeneity and its prothrombinase catalytic activity was examined. Time‐course conversion studies revealed that all prothrombin was converted to thrombin by purified hamster FGL2. Our biochemical studies demonstrate that FGL2 is a lipid‐activated serine protease and functions as a lectin by binding specific carbohydrate residues. Co‐immunoprecipitation analysis demonstrated that FGL2 of cauda epididymal fluid is ubiquitinated but not the FGL1. We propose that FGL2/FGL1 oligomers represent a novel and unique mechanism to shield the viable sperm population from degenerating spermatozoa contained within the tubule lumen.

中文翻译:

仓鼠尾附睾液的纤维蛋白原相关蛋白 FGL2:其凝血酶原酶活性的纯化、动力学分析及其在分离无活力精子中的作用

尽管附睾环境促进了精子的成熟和存活,但并非所有精子在通过附睾的过程中都能保持活力。附睾是否具有将活精子与有缺陷的精子分离的保护机制?以前,我们确定了 260/280 kDa 寡聚体(称为 eFGL-附睾纤维蛋白原样寡聚体)由两个二硫键连接的亚基组成:一个 64 kDa 的多肽被鉴定为纤维蛋白原样蛋白 2 (FGL2),一个 33 kDa 的多肽被鉴定为纤维蛋白原样蛋白-1 (FGL1)。我们的形态学研究表明,从附睾尾的主要细胞分泌的 eFGL 聚合成死亡茧状复合物(DCF),掩盖了有缺陷的管腔精子,但不能掩盖有活力的精子群体。在目前的研究中,我们从仓鼠附睾液中纯化 FGL2 使其趋于均匀,并检查了其凝血酶原酶催化活性。时程转化研究表明,所有的凝血酶原都被纯化的仓鼠 FGL2 转化为凝血酶。我们的生化研究表明,FGL2 是一种脂质激活的丝氨酸蛋白酶,通过结合特定的碳水化合物残基起到凝集素的作用。免疫共沉淀分析表明,附睾尾液的 FGL2 是泛素化的,但 FGL1 不是。我们建议 FGL2/FGL1 寡聚体代表了一种新颖独特的机制,可以保护活精子群免受小管腔内包含的退化精子的影响。时程转化研究表明,所有的凝血酶原都被纯化的仓鼠 FGL2 转化为凝血酶。我们的生化研究表明,FGL2 是一种脂质激活的丝氨酸蛋白酶,通过结合特定的碳水化合物残基起到凝集素的作用。免疫共沉淀分析表明,附睾尾液的 FGL2 是泛素化的,但 FGL1 不是。我们建议 FGL2/FGL1 寡聚体代表了一种新颖独特的机制,可以保护活精子群免受小管腔内包含的退化精子的影响。时程转化研究表明,所有的凝血酶原都被纯化的仓鼠 FGL2 转化为凝血酶。我们的生化研究表明,FGL2 是一种脂质激活的丝氨酸蛋白酶,通过结合特定的碳水化合物残基起到凝集素的作用。免疫共沉淀分析表明,附睾尾液的 FGL2 是泛素化的,但 FGL1 不是。我们建议 FGL2/FGL1 寡聚体代表了一种新颖独特的机制,可以保护活精子群免受小管腔内包含的退化精子的影响。
更新日期:2020-12-30
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