Lehmann Roland, Melle Christian, Escher Niko, von Eggeling Ferdinand
Core Unit Chip Application (CUCA), Institute of Human Genetics and Anthropology, Friedrich-Schiller-University, 07740 Jena, Germany.
J Proteome Res. 2005 Sep-Oct;4(5):1717-21. doi: 10.1021/pr050163s.
The aim of this work was to establish an approach for identification of protein interactions. This assay used an anti-S100A8 antibody coupled on beads and incubated with cell extract. The bead eluates were analyzed using ProteinChip technology and subsequently subjected to an appropriate digestion. Molecular masses of digestion fragments were determined by SELDI-MS, and database analysis revealed S100A10 as interacting protein. This result was confirmed by co-immunoprecipitation and immunocapturing. Using S100A10 as new bait, a specific interaction with S100A7 was detectable.
这项工作的目的是建立一种鉴定蛋白质相互作用的方法。该检测使用偶联在磁珠上的抗S100A8抗体,并与细胞提取物一起孵育。使用蛋白质芯片技术分析磁珠洗脱液,随后进行适当的消化。通过表面增强激光解吸电离飞行时间质谱(SELDI-MS)测定消化片段的分子量,数据库分析显示S100A10为相互作用蛋白。通过免疫共沉淀和免疫捕获证实了这一结果。以S100A10作为新的诱饵,可检测到与S100A7的特异性相互作用。