Xu Daosong, Hemler Martin E
Dana-Farber Cancer Institute and Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Mol Cell Proteomics. 2005 Aug;4(8):1061-71. doi: 10.1074/mcp.M400207-MCP200. Epub 2005 May 18.
Cell surface CD147 protein promotes production of matrix metalloproteinases and hyaluronan, associates with monocarboxylate transporters and integrins, and is involved in reproductive, neural, inflammatory, and tumor functions. Here we combined covalent cross-linking, mass spectrometric protein identification, and co-immunoprecipitation to show selective CD147 association with three major types of transporters (CD98 heavy chain (CD98hc)-L-type amino acid transporter, ASCT2, and monocarboxylate transporters) as well as a regulator of cell proliferation (epithelial cell adhesion molecule). In the assembly of these multicomponent complexes, CD147 and CD98hc play a central organizing role. RNA interference knock-down experiments established a strong connection between CD147 and CD98hc expression and a strong positive association of CD147 (and CD98hc) with cell proliferation. As the CD147-CD98hc complex and proliferation diminished, AMP-activated protein kinase (a cellular "fuel gauge") became activated, indicating a disturbance of cellular energy metabolism. Our data point to a CD147-CD98 cell surface supercomplex that plays a critical role in energy metabolism, likely by coordinating transport of lactate and amino acids. Furthermore we showed how covalent cross-linking, together with mass spectrometry, can be used to identify closely associated transmembrane proteins. This approach should also be applicable to many other types of transmembrane proteins besides those associated with CD98hc and CD147.
细胞表面的CD147蛋白可促进基质金属蛋白酶和透明质酸的产生,与单羧酸转运蛋白和整合素相关联,并参与生殖、神经、炎症和肿瘤功能。在此,我们结合了共价交联、质谱蛋白质鉴定和免疫共沉淀技术,以显示CD147与三种主要类型的转运蛋白(CD98重链(CD98hc)-L型氨基酸转运体、ASCT2和单羧酸转运蛋白)以及细胞增殖调节因子(上皮细胞粘附分子)的选择性结合。在这些多组分复合物的组装过程中,CD147和CD98hc发挥着核心组织作用。RNA干扰敲低实验证实了CD147和CD98hc表达之间的紧密联系,以及CD147(和CD98hc)与细胞增殖之间的强正相关。随着CD147-CD98hc复合物和细胞增殖的减少,AMP激活的蛋白激酶(一种细胞“燃料表”)被激活,表明细胞能量代谢受到干扰。我们的数据表明,CD147-CD98细胞表面超复合物可能通过协调乳酸和氨基酸的转运在能量代谢中起关键作用。此外,我们展示了共价交联与质谱联用如何用于鉴定紧密相关的跨膜蛋白。这种方法除了适用于与CD98hc和CD147相关的跨膜蛋白外,还应适用于许多其他类型的跨膜蛋白。