Reconstitution of peptide-binding activity by TAP in proteoliposomes.
The purification and functional reconstitution of the transporter associated with antigen processing (TAP) is crucial for a complete molecular understanding of its action. Here, we report the conditions for the successful solubilization of human TAP from cellular membranes while maintaining TAP peptide-binding activity. In addition, solubilized TAP was incorporated into proteoliposomes and shown to possess specific peptide-binding activity. These studies provide the foundation for future attempts to achieve the complete functional reconstitution of TAP, which includes peptide transport.
Stephens
D B
DB
H. Lee Moffitt Cancer Center and Research Institute, and the Department of Biochemistry and Molecular Biology, University of South Florida College of Medicine, Tampa 33612, USA.
Androlewicz
M J
MJ
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000378:metabolism*; D000595:Amino Acid Sequence; D001665:Binding Sites; D002051:Burkitt Lymphoma; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D008081:Liposomes; D009842:Oligopeptides / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry; D011510:Proteolipids / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D018528:ATP-Binding Cassette Transporters; D008081:Liposomes; D009842:Oligopeptides; D010743:Phospholipids; D011510:Proteolipids; C020485:proteoliposomes