McGee R S, Herr J C
Department of Anatomy and Cell Biology, University of Virginia Medical Center, Charlottesville 22908.
Biol Reprod. 1988 Sep;39(2):499-510. doi: 10.1095/biolreprod39.2.499.
Human seminal vesicle and prostatic fluids were obtained separately and reconstituted in vitro to test the hypothesis that proteolytic enzymes of prostatic origin would degrade seminal vesicle-specific antigen (SVSA). Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis to follow the fate of SVSA over time, we found that upon mixing the two secretions, SVSA was converted to forms of intermediate and low molecular weight identical to transformations seen in normal liquefied ejaculates. Diisoproprylfluorophophate, a serine protease inhibitor, prevented this degradation, indicating serine protease involvement in the proteolysis of SVSA. Prostate-specific antigen (PSA; also known as P-30), recently identified as a serine protease, was examined for its ability to mimic the effects of prostatic fluid on SVSA. Purified PSA catalyzed degradation of SVSA to produce proteolytic fragments that comigrated and were immunologically related to SVSA fragments produced by prostatic fluid. Purified PSA in the presence of serine protease inhibitors was unable to degrade SVSA. These results demonstrate that SVSA is a substrate for PSA during human semen liquefaction.
分别获取人精囊液和前列腺液，并在体外进行重组，以验证前列腺来源的蛋白水解酶会降解精囊特异性抗原（SVSA）这一假说。利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹分析来追踪SVSA随时间的变化情况，我们发现，将两种分泌物混合后，SVSA会转化为与正常液化精液中所见转化形式相同的中低分子量形式。丝氨酸蛋白酶抑制剂二异丙基氟磷酸可阻止这种降解，表明丝氨酸蛋白酶参与了SVSA的蛋白水解过程。最近被鉴定为丝氨酸蛋白酶的前列腺特异性抗原（PSA；也称为P-30），被检测其模拟前列腺液对SVSA影响的能力。纯化的PSA催化SVSA的降解，产生的蛋白水解片段与前列腺液产生的SVSA片段迁移一致且具有免疫相关性。在丝氨酸蛋白酶抑制剂存在的情况下，纯化的PSA无法降解SVSA。这些结果表明，在人类精液液化过程中，SVSA是PSA的底物。
