Department of Medical Biochemistry, Shiga University of Medical Science, Seta, Otsu 520-2192.
J Biol Chem. 2010 Jan 15;285(3):2184-92. doi: 10.1074/jbc.M109.063495. Epub 2009 Nov 17.
We found that factor H (FH) exists in porcine seminal plasma. Purified FH strongly inhibited serum alternative pathway complement activation against lipopolysaccharide. The molecular weight, pI, and heparin-binding activity of the purified protein were different from those of purified FH from porcine serum. The complement regulatory activity of seminal plasma FH was approximately 2-fold stronger than that of serum FH. Treatment of purified serum FH with sialidase and N-glycosidase F gave almost the same results as those of seminal plasma FH. The deletion of sialic acid from the carbohydrate chains of both FHs contributed to heparin-binding and complement regulatory activities. Results of reverse transcriptase-PCR, Western blot analysis, and immunohistochemistry showed that seminal plasma FH is mainly secreted from epithelial cells of the seminal vesicle in male genital tracts. FH was also detected in the outer acrosomal region of ejaculated sperm by immunofluorescence staining, and found that the purified FH from the sperm membrane has the same complement regulatory activity as that of seminal plasma FH. The ejaculated sperm possessing FH in the outer acrosomal region considerably evaded complement attack. We also found that there is strong complement activity in fluids from female genital tract ducts. These findings indicate that FH bound to the outer acrosomal region and soluble FH play important roles in protecting sperm against complement attack in male and female genital tracts.
我们发现因子 H(FH)存在于猪的精液中。纯化的 FH 强烈抑制血清替代途径补体对脂多糖的激活。该纯化蛋白的分子量、等电点和肝素结合活性与来自猪血清的纯化 FH 不同。精液 FH 的补体调节活性比血清 FH 强约 2 倍。唾液酸酶和 N-糖苷酶 F 处理纯化的血清 FH 得到的结果与精液 FH 几乎相同。两种 FH 的糖链上唾液酸的缺失有助于肝素结合和补体调节活性。逆转录 PCR、Western blot 分析和免疫组织化学的结果表明,精液 FH 主要由生殖道精囊的上皮细胞分泌。免疫荧光染色还在射出精子的外顶体区检测到 FH,并且发现从精子膜中纯化的 FH 具有与精液 FH 相同的补体调节活性。在外顶体区具有 FH 的射出精子能显著逃避补体攻击。我们还发现女性生殖道管腔内存在强烈的补体活性。这些发现表明,结合在外顶体区的 FH 和可溶性 FH 在保护精子免受生殖道内补体攻击方面发挥着重要作用。