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ATP-Binding Cassette Transporter B4 Anchors the Cell Adhesion Molecule DdCAD-1 to Cell Membrane in Dictyostelium discoideum
Authors:Chunxia Yang  Liansheng Hou  Qixiu Yang  Chi-Hung Siu
Institution:2. School of Life Science, East China Normal University, 200062, Shanghai, China
1. Banting and Best Department of Medical Research and the Department of Biochemistry, University of Toronto, Toronto, Ontario, M5S 1A8, Canada
Abstract: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.
Keywords:Anchoring protein  Co-immunoprecipitation  Cross-linking  DdCAD-1  ABC transporters
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