Rasmussen U B, Wohlrab H
Biochim Biophys Acta. 1986 Dec 3;852(2-3):306-14. doi: 10.1016/0005-2728(86)90236-7.
Peptide maps were generated of the CNBr-digested mitochondrial phosphate-transport protein and ADP/ATP carrier from bovine and rat heart, rat liver and blowfly flight muscle. Total mitochondrial proteins from the same sources plus pig heart were separated by SDS-polyacrylamide gel electrophoresis. The peptide maps and the total mitochondrial proteins were electroblotted onto nitrocellulose membranes and reacted with rabbit antisera raised against the purified bovine heart phosphate-transport protein and the ADP/ATP carrier. On the basis of antibody specificity, mobility in SDS-polyacrylamide gel electrophoresis, and peptide maps the following was concluded. Phosphate-transport protein alpha and phosphate-transport protein beta (pig and bovine heart) react equally with the first and also with the second of two independent phosphate-transport protein-antisera. Tissue-specific structural domains exist for both the phosphate-transport protein and the ADP/ATP carrier, i.e., one phosphate-transport protein-antiserum reacts with the phosphate-transport protein from all assayed sources, the other only with the cardiac phosphate-transport protein. These differences may reflect tissue-specific regulation of phosphate and adenine nucleotide transport. Homologies among the different species are found for the phosphate transport protein and the ADP/ATP carrier, except for the flight muscle ADP/ATP carrier. These conserved structural domains of the phosphate-transport protein may relate directly to catalytic activity. Alkylation of the purified phosphate-transport proteins and the ADP/ATP carriers by the transport inhibitor N-ethylmaleimide affects electrophoretic mobilities but not the antibody binding. Neither of the two phosphate-transport protein-antisera nor the ADP/ATP-carrier antiserum react with both phosphate transport protein and ADP/ATP carrier, even though these two proteins possess similarities in primary structure and function. Possible mechanisms for generating tissue-specific structural differences in the proteins are discussed.
生成了来自牛和大鼠心脏、大鼠肝脏及家蝇飞行肌的经溴化氰消化的线粒体磷酸转运蛋白和 ADP/ATP 载体的肽图。来自相同来源以及猪心脏的总线粒体蛋白通过 SDS - 聚丙烯酰胺凝胶电泳进行分离。将肽图和总线粒体蛋白电印迹到硝酸纤维素膜上，并与针对纯化的牛心脏磷酸转运蛋白和 ADP/ATP 载体产生的兔抗血清反应。基于抗体特异性、在 SDS - 聚丙烯酰胺凝胶电泳中的迁移率以及肽图，得出以下结论。磷酸转运蛋白α和磷酸转运蛋白β（猪和牛心脏）与两种独立的磷酸转运蛋白抗血清中的第一种以及第二种反应程度相同。磷酸转运蛋白和 ADP/ATP 载体均存在组织特异性结构域，即一种磷酸转运蛋白抗血清与所有检测来源的磷酸转运蛋白反应，另一种仅与心脏磷酸转运蛋白反应。这些差异可能反映了磷酸盐和腺嘌呤核苷酸转运的组织特异性调节。除了飞行肌 ADP/ATP 载体外，在不同物种间发现了磷酸转运蛋白和 ADP/ATP 载体的同源性。磷酸转运蛋白的这些保守结构域可能与催化活性直接相关。转运抑制剂 N - 乙基马来酰亚胺对纯化的磷酸转运蛋白和 ADP/ATP 载体的烷基化作用影响电泳迁移率，但不影响抗体结合。两种磷酸转运蛋白抗血清和 ADP/ATP 载体抗血清均不与磷酸转运蛋白和 ADP/ATP 载体同时反应，尽管这两种蛋白在一级结构和功能上具有相似性。文中讨论了在这些蛋白中产生组织特异性结构差异的可能机制。
