Mariottini P, Chomyn A, Attardi G, Trovato D, Strong D D, Doolittle R F
Cell. 1983 Apr;32(4):1269-77. doi: 10.1016/0092-8674(83)90308-2.
Antibodies prepared against chemically synthesized peptides predicted from the DNA sequence have been used to detect human mitochondrial gene products. In particular, antibodies directed against either the NH2-terminal decapeptide or the COOH-terminal undecapeptide of cytochrome c oxidase subunit II (COII) were both very effective in immunoprecipitating the previously identified COII polypeptide from an SDS lysate of mitochondria from HeLa cells. Similarly, antibodies directed against the COOH-terminal nonapeptide of the putative polypeptide encoded in the unidentified reading frame A6L, which overlaps the ATPase 6 gene, immunoprecipitated specifically a component (#25) of the HeLa cell mitochondrial translation products; antibodies directed against the NH2-terminal octapeptide also precipitated protein 25, although less efficiently. The size of protein 25, as estimated from its electrophoretic mobility, is compatible with its being the unidentified reading frame A6L product. Furthermore, a fingerprinting analysis of this protein after trypsin digestion has given results consistent with this identification.
针对根据DNA序列预测的化学合成肽制备的抗体已用于检测人类线粒体基因产物。特别是，针对细胞色素c氧化酶亚基II（COII）的NH2末端十肽或COOH末端十一肽的抗体，在从HeLa细胞线粒体的SDS裂解物中免疫沉淀先前鉴定的COII多肽方面都非常有效。同样，针对与ATPase 6基因重叠的未鉴定阅读框A6L中编码的假定多肽的COOH末端九肽的抗体，特异性免疫沉淀了HeLa细胞线粒体翻译产物的一种成分（#25）；针对NH2末端八肽的抗体也沉淀了蛋白质25，尽管效率较低。根据其电泳迁移率估计，蛋白质25的大小与其作为未鉴定阅读框A6L产物相符。此外，对该蛋白质进行胰蛋白酶消化后的指纹分析结果与该鉴定一致。
