Characterization of a membrane antigen from rabbit testis and sperm isolated by using monoclonal antibodies and effect of its antiserum on fertility.
Characterization of a membrane antigen from rabbit testis and sperm isolated by using monoclonal antibodies and effect of its antiserum on fertility.
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使用单克隆抗体分离的兔睾丸和精子膜抗原的表征及其抗血清对生育力的影响。
DOI：
10.1073/pnas.81.3.857
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发表时间：
1984
影响因子：
11.1
通讯作者：
Menge,AC
中科院分区：
文献类型：
--
作者：
Naz,RK;Rosenblum,BB;Menge,AC
用含有抗生育力单克隆抗体（8C10.5）IgG的免疫吸附柱，从脱氧胆酸盐溶解的兔睾丸和精子中分离抗原。通过逐步增加pH（8.0、10.0和11.4）进行洗脱，其中pH 11.4的级分在通过柱再循环后，在用银染色的平板NaDodSO 4/PAGE中显示63千道尔顿的单一条带。抗原分子由两个亚基组成，在二维PAGE上显示出相同分子大小范围（50-70千道尔顿）内但电荷不同的许多斑点。从睾丸和精子中分离的抗原主要显示相同的斑点。抗原为高碘酸/希夫氏阳性，含有21%的碳水化合物。抗原的去唾液酸衍生物在NaDodSO 4/PAGE上不改变其特征。这种糖蛋白分解成两种类型的多肽，那些结合和那些不结合到一个透镜culinaris凝集素柱;一些多肽出现共同的两个馏分。鼠抗血清对抗原既不凝集也不固定的兔精子，但在免疫荧光反应与质膜的活兔精子，以及与小鼠和人的精子。用处理过的精子授精的雌兔的受精没有受到影响，但是，到第9天，生育力显著降低（对照组的21%）。受精后的抗生育作用不是由于孤雌激活或多精受精。抗血清在一维和二维凝胶电泳转移印迹过程中与一个特定的条带反应，并且不受不同体细胞组织吸收的影响。
An antigen was isolated from deoxycholate-solubilized rabbit testis and sperm by using an immunosorbent column containing IgG from a monoclonal antibody (8C10.5) that inhibits fertility. Elution was by stepwise increases in pH (8.0, 10.0, and 11.4), with the pH 11.4 fraction after recycling through the column showing a single band at 63 kilodaltons in slab NaDodSO4/PAGE with a silver stain. The antigen molecule was composed of two subunits, which on two-dimensional PAGE showed many spots within the same molecular size range (50-70 kilodaltons) but differing in charge. The antigen isolated either from testis or sperm showed mainly the same spots. The antigen is periodic acid/Schiff positive and contained 21% carbohydrate. An asialo-derivative of the antigen did not change its characteristics on NaDodSO4/PAGE. This glycoprotein resolved into two types of polypeptides, those binding and those not binding to a lens culinaris lectin column; some of the polypeptides appeared common to both fractions. Murine antiserum against the antigen neither agglutinated nor immobilized rabbit sperm but in immunofluorescence reacted with the plasma membrane of viable rabbit sperm as well as with murine and human sperm. Fertilization of female rabbits inseminated with treated sperm was not affected, but, by 9 days, fertility was significantly reduced (21% of controls). The postfertilization antifertility effect was not due to parthenogenic activation or to polyspermy. The antiserum reacted with one specific band in the one- and two-dimensional gel electrophoretic transfer blot procedure and was unaffected by absorption with different somatic tissues.