Sebald W, Machleidt W, Wachter E
Proc Natl Acad Sci U S A. 1980 Feb;77(2):785-9. doi: 10.1073/pnas.77.2.785.
The N,N'-dicyclohexylcarbodiimide-binding proteolipid subunit of the mitochondrial adenosinetriphosphatases (ATP phosphohydrolase, EC 3.6.1.3) of Neurospora crassa and Saccharomyces cerevisiae were purified from mitochondria incubated with the radioactively labeled inhibitor. The specifically labeled subunit was cleaved with cyanogen bromide and N-bromosuccinimide, and the resultant fragments were separated by gel chromatography in the presence of 80% (vol/vol) formic acid. The N,N'-dicyclohexylcarbodiimide label was recovered in each organism exclusively in a 17-residue fragment. Further analysis by automated solid-phase Edman degradation revealed that the bound label was present at only one position, corresponding to a glutamyl residue. The N,N'-dicyclohexylcarbodiimide-modified glutamyl residue is the only identical acidic position in both proteins and occurs in the middle of a hydrophobic sequence of about 25 residues.
从与放射性标记抑制剂一起孵育的线粒体中纯化了粗糙脉孢菌和酿酒酵母线粒体腺苷三磷酸酶（ATP磷酸水解酶，EC 3.6.1.3）的N，N'-二环己基碳二亚胺结合蛋白脂质亚基。用溴化氰和N-溴代琥珀酰亚胺切割特异性标记的亚基，并在80%（体积/体积）甲酸存在下通过凝胶色谱法分离所得片段。在每种生物体中，N，N'-二环己基碳二亚胺标记仅在一个17个残基的片段中回收。通过自动固相埃德曼降解进一步分析表明，结合的标记仅存在于一个位置，对应于一个谷氨酰残基。N，N'-二环己基碳二亚胺修饰的谷氨酰残基是两种蛋白质中唯一相同的酸性位置，且位于约25个残基的疏水序列中间。
