Richardson R, Buckingham T, Boettger H, Poirier G R
J Reprod Immunol. 1987 Jun;11(2):101-16. doi: 10.1016/0165-0378(87)90015-5.
A monoclonal antibody, J-23, was produced to an epitope of a binding (acceptor) component on the plasma membrane in the acrosomal cap region of the mouse sperm head. The component binds a proteinase inhibitor of seminal vesicle origin at ejaculation and participates in the in vitro binding of capacitated sperm to the zona pellucida. The antibody, an IgM molecule, recognizes affinity purified acceptor, crude acceptor and whole sperm as determined by ELISA methodology. The antibody reacts with a 15,000 molecular weight component, the size previously determined for the acceptor, found in the supernatants of frozen-thawed cauda epididymal sperm. In addition, it binds to a 21,000 molecular weight component generated by mixing an excess of purified inhibitor (6400 daltons) with a crude acceptor preparation. J-23 binds to an epitope in the same region of the sperm head as does the inhibitor. This epitope becomes fully expressed during epididymal maturation and is found only in the lumen of epididymal tissues. Pretreating sperm with J-23 inhibits their ability to bind the inhibitor as well as the zona pellucida. Pretreating sperm with the inhibitor has little effect on the binding of J-23. These data indicate that J-23 recognizes an epitope on the acceptor but the epitope is not directly involved with inhibitor binding.
一种单克隆抗体J-23是针对小鼠精子头部顶体帽区域质膜上结合(受体)成分的一个表位产生的。该成分在射精时结合精囊来源的蛋白酶抑制剂,并参与获能精子与透明带的体外结合。通过酶联免疫吸附测定法(ELISA)确定,该抗体(一种IgM分子)能识别亲和纯化的受体、粗提受体和完整精子。该抗体与分子量为15000的成分发生反应,这是先前确定的受体的大小,在冻融附睾尾部精子的上清液中可以找到。此外,它还能与通过将过量纯化抑制剂(6400道尔顿)与粗提受体制剂混合产生的分子量为21000的成分结合。J-23与抑制剂一样,结合精子头部的同一区域的一个表位。这个表位在附睾成熟过程中完全表达,并且只存在于附睾组织的管腔中。用J-23预处理精子会抑制它们结合抑制剂以及透明带的能力。用抑制剂预处理精子对J-23的结合影响很小。这些数据表明,J-23识别受体上的一个表位,但该表位不直接参与抑制剂的结合。