School of Life Science, East China Normal University, 200062 Shanghai, China.
Banting and Best Department of Medical Research and the Department of Biochemistry, University of Toronto, Toronto, Ontario M5S 1A8 Canada.
Indian J Microbiol. 2013 Dec;53(4):460-6. doi: 10.1007/s12088-013-0393-0. Epub 2013 Mar 22.
In Dictyostelium, soluble cell adhesion molecule, DdCAD-1, regulates cell-cell interaction through an unknown anchoring protein on the plasma membrane. Far western blot analysis using different probes revealed that the potential DdCAD-1 interacting protein was between 64 and 98 kDa. To isolate and identify the anchoring protein, GST-DdCAD-1 and anchoring protein were cross-linked in vivo by chemical cross-linker and stable protein complex was isolated by co-immunoprecipitation assays. The protein cross-linked to DdCAD-1 was extracted from the gel slice and trypsinized. The peptides were subjected to analysis by mass spectrometry, which showed that the putative anchoring protein belongs to ATP-binding cassette transporter family.
在粘菌中，可溶性细胞黏附分子 DdCAD-1 通过质膜上未知的锚定蛋白调节细胞间相互作用。使用不同探针的 Far western blot 分析表明，潜在的 DdCAD-1 相互作用蛋白的大小在 64 和 98 kDa 之间。为了分离和鉴定锚定蛋白，使用化学交联剂在体内交联 GST-DdCAD-1 和锚定蛋白，并通过共免疫沉淀测定法分离稳定的蛋白复合物。从凝胶片上提取与 DdCAD-1 交联的蛋白，并进行胰蛋白酶消化。肽段通过质谱分析，表明该假定的锚定蛋白属于 ATP 结合盒转运蛋白家族。
