Functional cysteine-less subunits of the transporter associated with antigen processing (TAP1 and TAP2) by de novo gene assembly.
Within the adaptive immune system the transporter associated with antigen processing (TAP) plays a pivotal role in loading of peptides onto major histocompatibility (MHC) class I molecules. As a central tool to investigate the structure and function of the TAP complex, we created cysteine-less human TAP subunits by de novo gene synthesis, replacing all 19 cysteines in TAP1 and TAP2. After expression in TAP-deficient human fibroblasts, cysteine-less TAP1 and TAP2 are functional with respect to adenosine triphosphate (ATP)-dependent peptide transport and inhibition by ICP47 from herpes simplex virus. Cysteine-less TAP1 and TAP2 restore maturation and intracellular trafficking of MHC class I molecules to the cell surface.
Heintke
Susanne
S
Institut für Biochemie, Biozentrum, Johann Wolfgang Goethe-Universität Frankfurt, Marie Curie Str. 9, D-60439, Frankfurt/M, Germany.
Chen
Min
M
Ritz
Ulrike
U
Lankat-Buttgereit
Brigitte
B
Koch
Joachim
J
Abele
Rupert
R
Seliger
Barbara
B
Tampé
Robert
R
D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D000071450:ATP Binding Cassette Transporter, Subfamily B, Member 3; D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D019943:Amino Acid Substitution; D017951:Antigen Presentation; D001693:Biological Transport, Active; D002460:Cell Line; D003545:Cysteine / Q000737:chemistry; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D000071450:ATP Binding Cassette Transporter, Subfamily B, Member 3; D018528:ATP-Binding Cassette Transporters; D015395:Histocompatibility Antigens Class I; D021122:Protein Subunits; D011994:Recombinant Proteins; C073119:TAP1 protein, human; C071692:TAP2 protein, human; D000255:Adenosine Triphosphate; D003545:Cysteine