Remold-O'Donnell E, Kenney D M, Parkman R, Cairns L, Savage B, Rosen F S
J Exp Med. 1984 Jun 1;159(6):1705-23. doi: 10.1084/jem.159.6.1705.
gpL115 is a lymphocyte surface component that is deficient in patients with the X-chromosome-linked immune deficiency Wiskott-Aldrich syndrome (6). The glycoprotein nature of gpL115 is demonstrated through labeling in carbohydrate moieties by [3H]NaBH4 and its synthesis by lymphocytes through labeling with [35S]methionine. Native gpL115 adheres to wheat germ lectin-Sepharose and sialidase-treated gpL115 does not adhere, indicating that native gpL115 adheres via clusters of sialic acid residues. When tested on peanut lectin, which shows specificity for the disaccharide Gal beta 1-3GalNAc, gpL115 is nonadherent and sialidase-treated gpL115 is adherent, indicating the presence of the sequence sialic acid-Gal beta 1-3GalNAc, which is characteristic for O-linked (mucin-type, acidic-type) carbohydrates. A surface glycoprotein with all the above characteristics was found on the lymphoblastoid cell line CEM. CEM cells were used as immunogen to generate the monoclonal antibody L10, an IgG1, which binds native and sialidase-treated gpL115 . Sialidase-treatment of gpL115 significantly alters its physical properties, reducing its electrophoretic mobility and changing its behavior on isoelectrofocusing. Cumulatively, these findings indicate that gpL115 , like glycophorin of erythrocytes and GPIb of platelets, is a sialoglyco protein with significant quantities of O-linked carbohydrate. On treatment with limiting sialidase concentrations, gpL115 of normal lymphocytes is transformed into a series of partially desialylated species of decreasing electrophoretic mobility. This finding resembles the situation with lymphocytes of some Wiskott-Aldrich syndrome patients. Lymphocytes of eight Wiskott-Aldrich syndrome patients were found to be deficient in 125I-labeled gpL115 . Lymphocytes from three of these patients displayed an abnormal 125I-component of apparent mol wt 135,000.
gpL115是一种淋巴细胞表面成分,在患有X染色体连锁免疫缺陷威斯科特-奥尔德里奇综合征的患者中缺乏(6)。gpL115的糖蛋白性质通过用[3H]硼氢化钠标记碳水化合物部分得以证明,并且淋巴细胞通过用[35S]甲硫氨酸标记来合成它。天然的gpL115能与麦胚凝集素-琼脂糖结合,而经唾液酸酶处理的gpL115则不能结合,这表明天然的gpL115通过唾液酸残基簇进行结合。当用对二糖Galβ1-3GalNAc具有特异性的花生凝集素进行测试时,gpL115不结合,而经唾液酸酶处理的gpL115结合,这表明存在唾液酸-Galβ1-3GalNAc序列,这是O-连接(粘蛋白型、酸性型)碳水化合物的特征。在淋巴母细胞系CEM上发现了一种具有上述所有特征的表面糖蛋白。CEM细胞被用作免疫原以产生单克隆抗体L10,一种IgG1,它能结合天然的和经唾液酸酶处理的gpL115。对gpL115进行唾液酸酶处理会显著改变其物理性质,降低其电泳迁移率并改变其在等电聚焦时的行为。总的来说,这些发现表明gpL115与红细胞的血型糖蛋白和血小板的糖蛋白Ib一样,是一种含有大量O-连接碳水化合物的唾液酸糖蛋白。用有限浓度的唾液酸酶处理时,正常淋巴细胞的gpL115会转化为一系列电泳迁移率逐渐降低的部分去唾液酸化的物种。这一发现类似于一些威斯科特-奥尔德里奇综合征患者的淋巴细胞的情况。发现八名威斯科特-奥尔德里奇综合征患者的淋巴细胞缺乏125I标记的gpL115。其中三名患者的淋巴细胞显示出一种表观分子量为135,000的异常125I成分。