Generation of monoclonal antibodies to integrin-associated proteins - Evidence that alpha(3)beta(1) complexes with EMMPRIN/basigin/OX47/M6
Generation of monoclonal antibodies to integrin-associated proteins - Evidence that alpha(3)beta(1) complexes with EMMPRIN/basigin/OX47/M6
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DOI：
10.1074/jbc.272.46.29174
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发表时间：
1997-11-14
影响因子：
4.8
通讯作者：
Hemler, ME
中科院分区：
文献类型：
--
作者：
Berditchevski, F;Chang, S;Hemler, ME
α（3）β（1）整联蛋白与其他细胞表面蛋白形成复合物，包括跨膜-4超家族（TM 4SF）蛋白（例如CD 9、CD 53、CD 63、CD 81和CD 82）。为了鉴定与α（3）β（1）整联蛋白相关的其他细胞表面蛋白，开发了单克隆抗体选择方案。用从HT 1080纤维肉瘤细胞分离的含有整联蛋白α（3）β（1）的复合物免疫小鼠，然后产生712个杂交瘤克隆，并且95个分泌识别HT 1080细胞表面的抗体。除去了12种直接识别整合素α（3）或β（1）亚基的抗体。在剩余的83种中，16种在非严格洗涤剂条件下类似于整合素的蛋白质共免疫沉淀。这16个包括识别EMMPRIN/basigin/OX-47/M6的15个单克隆抗体，所述M6是具有两个免疫球蛋白结构域的45-55-kDa跨膜蛋白。如相互免疫沉淀实验所示，EMMPRIN蛋白与α（3）β（1）和α（6）β（1）相关，但不与α（2）β（1）或α（5）β（1）相关。此外，通过细胞表面交联和免疫荧光共定位实验证实了与α（3）β（1）的相关。重要的是，EMMPRIN-alpha（3）beta（1）复合物似乎不含TM 4SF蛋白，表明它们与TM 4SF蛋白-alpha（3）beta（1）复合物不同。
The alpha(3) beta(1) integrin forms complexes with other cell-surface proteins, including transmembrane-4 superfamily (TM4SF) proteins (e.g. CD9, CD53, CD63, CD81, and CD82). To identify additional cell-surface proteins associated with alpha(3) beta(1) integrin, a monoclonal antibody selection protocol was developed, Mice were immunized with integrin alpha(3) beta(1)-containing complexes isolated from HT1080 fibrosarcoma cells, and then 712 hybridoma clones were produced, and 95 secreted antibodies that recognized the HT1080 cell surface, Among these, 12 antibodies directly recognizing integrin alpha(3) or beta(1) subunits were eliminated, Of the remaining 83, 16 co-immunoprecipitated proteins that resembled integrins under non-stringent detergent conditions. These 16 included 15 monoclonal antibodies recognizing EMMPRIN/basigin/OX-47/M6, a 45-55-kDa transmembrane protein with two immunoglobulin domains. The EMMPRIN protein associated with alpha(3) beta(1) and alpha(6) beta(1), but not alpha(2) beta(1) or alpha(5) beta(1), as shown by reciprocal immunoprecipitation experiments, Also, association with alpha(3) beta(1) was confirmed by cell-surface cross-linking and immunofluorescence co-localization experiments, Importantly, EMMPRIN-alpha(3) beta(1) complexes appear not to contain TM4SF proteins, suggesting that they are distinct from TM4SF protein-alpha(3) beta(1) complexes.