Rönkkö S, Linnala-Kankkunen A, Tuhkanen A L
Department of Anatomy, University of Kuopio, Finland.
Andrologia. 1994 Mar-Apr;26(2):73-8. doi: 10.1111/j.1439-0272.1994.tb00760.x.
A fraction from bovine seminal vesicle fluid that initiated acrosome reaction of bovine epididymal spermatozoa in vitro in the presence of heparin was prepared by sequential chromatographies on heparin-Sepharose, gel filtration (Superose 12) and reversed phase chromatography (ProRPC). Sequence analysis of the separated fraction showed that it contained the major protein (PDC-109) with 100% homology. This fraction contained also Ca(2+)-dependent phospholipase A2-like activity which hydrolysed phosphatidylethanolamine and phosphatidylcholine with 14C-labelled linoleic (lino-PE, lino-PC) or arachidonic acid (ara-PE, ara-PC) at sn-2 position. This protein was not detected in N-terminal sequence analysis. Lysophosphatidylcholine, lysophosphatidylethanolamine, and p-bromophenacyl bromide (p-BPB) inhibited this lipolytic activity. Sulfoglycolipid (Seminolipid) had inhibitory effect at concentrations above 0.1 mM but activated slightly the enzyme at lower concentrations. Boiling destroyed acrosome initiating activity in the separated fraction.
通过在肝素 - 琼脂糖、凝胶过滤(Superose 12)和反相色谱(ProRPC)上进行连续色谱法,制备了来自牛精囊液的一种组分,该组分在肝素存在下可在体外引发牛附睾精子的顶体反应。对分离出的组分进行序列分析表明,它含有具有100%同源性的主要蛋白质(PDC - 109)。该组分还具有钙依赖性磷脂酶A2样活性,可在sn - 2位水解带有14C标记亚油酸(亚油酰 - 磷脂酰乙醇胺、亚油酰 - 磷脂酰胆碱)或花生四烯酸(花生四烯酰 - 磷脂酰乙醇胺、花生四烯酰 - 磷脂酰胆碱)的磷脂酰乙醇胺和磷脂酰胆碱。在N端序列分析中未检测到这种蛋白质。溶血磷脂酰胆碱、溶血磷脂酰乙醇胺和对溴苯甲酰溴(p - BPB)抑制这种脂解活性。硫糖脂(半乳糖脑苷脂)在浓度高于0.1 mM时具有抑制作用,但在较低浓度时可轻微激活该酶。煮沸会破坏分离组分中的顶体引发活性。