Functional cysteine-less subunits of the transporter associated with antigen processing (TAP1 and TAP2) by de novo gene assembly |
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Authors: | Heintke Susanne Chen Min Ritz Ulrike Lankat-Buttgereit Brigitte Koch Joachim Abele Rupert Seliger Barbara Tampé Robert |
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Affiliation: | Institute of Physiology and Centre for Integrated Genomics, Academy of Sciences of the Czech Republic, 142 20 4 - Krc, Prague, Czech Republic. |
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Abstract: | 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. |
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