The light chain of CD98 is identified as E16/TA1 protein
The light chain of CD98 is identified as E16/TA1 protein
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DOI：
10.1074/jbc.273.50.33127
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发表时间：
1998-12-11
影响因子：
4.8
通讯作者：
Hemler, ME
中科院分区：
文献类型：
--
作者：
Mannion, BA;Kolesnikova, TV;Hemler, ME
80/40-kDa CD 98蛋白。使用偶联到琼脂糖珠的抗CD 98重链单克隆抗体纯化复合物。将开槽的蛋白质进行制备SDS-聚丙烯酰胺凝胶电泳，切下对应于40-kDa CD 98轻链的蛋白质。在用胰蛋白酶进行蛋白水解后，通过质谱法对肽片段进行测序。获得的9个残基与人E16和大鼠TAI蛋白的已建立的C-末端序列相同，表明TA 1/E16蛋白是CD 98轻链。与此一致，抗TA 1/E16抗体特异性免疫印迹类似于人CD 98复合物免疫沉淀时存在的35-40-kDa轻链。此外，抗CD 98重链抗体特异性地免疫共沉淀来自用血凝素标记的E16 cDNA转染的细胞的血凝素标记的轻链。总之，基于部分氨基酸序列同一性、抗体交叉反应性、遗传重建证据、相似的分子大小和可比的细胞分布，CD 98轻链与TA 1/E16蛋白相同。
The 80/40-kDa CD98 protein. complex was purified using an anti-CD98 heavy chain monoclonal antibody coupled to Sepharose beads. fluted proteins were subjected to preparative SDS-polyacrylamide gel electrophoresis, and protein corresponding to the 40-kDa CD98 light chain was excised. Following proteolysis with trypsin, a peptide fragment was sequenced by mass spectrometry. The nine residues obtained we:re identical to established C-terminal sequences of the human E16 and rat TAI proteins, suggesting that TA1/E16 protein is the CD98 Light chain. Consistent with this, anti-TA1/E16 antibodies specifically immunoblotted the similar to 35-40-kDa light chain present upon immunoprecipitation of the human CD98 complex. Furthermore, anti-CD98 heavy chain antibody specifically co-immunoprecipitated hemagglutinin-tagged light chain from cells transfected with hemagglutinin-tagged E16 cDNA. In conclusion, the CD98 light chain is identical to the TA1/E16 protein, based on partial amino acid sequence identity, antibody crossreactivity, genetic reconstitution evidence, similar molecular size, and comparable cell distribution.