Functional cysteine‐less subunits of the transporter associated with antigen processing (TAP1 and TAP2) by de novo gene assembly

FEBS Letters

Abele, Rupert¹  Tampé, Robert¹  Heintke, Susanne¹  Lankat-Buttgereit, Brigitte²  Chen, Min¹  Seliger, Barbara³  Koch, Joachim¹  Ritz, Ulrike³ 

[1] Institut für Biochemie, Biozentrum, Johann Wolfgang Goethe-Universität Frankfurt, Marie Curie Str. 9, D-60439 Frankfurt/M, Germany;Institut für Physiologische Chemie, Philipps-Universität Marburg, Karl-von-Frisch-Str. 1, D-35043 Marburg, Germany;III. Med. Klinik and Poliklinik, Johannes Gutenberg-Universität Klinikum Mainz, Langenbeckstr. 1, D-55131 Mainz, Germany

关键词: Adenosine triphosphate-binding cassette transporter;    Antigen processing;    Cysteine-scanning mutagenesis;    Membrane protein;    ECL;    enhanced chemiluminescence;    FACS;    fluorescence-activated cell sorting;    MHC;    major histocompatibility complex;    NBD;    nucleotide-binding domain;    TAP;    transporter associated with antigen processing;    TMD;    transmembrane domain;   

DOI  :  10.1016/S0014-5793(02)03746-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown