TY  - JOUR
A1  - Schölz, Christian
A1  - Parcej, David
A1  - Stenby Ejsing, Christer
A1  - Robenek, Horst
A1  - Urbatsch, Ina L.
A1  - Tampé, Robert
T1  - Specific lipids modulate the transporter associated with antigen processing (TAP)
T2  - Journal of biological chemistry
N2  - The transporter associated with antigen processing (TAP) plays a key role in adaptive immunity by translocating proteasomal degradation products from the cytosol into the endoplasmic reticulum lumen for subsequent loading onto major histocompatibility (MHC) class I molecules. For functional and structural analysis of this ATP-binding cassette complex, we established the overexpression of TAP in the methylotrophic yeast Pichia pastoris. Screening of optimal solubilization and purification conditions allowed the isolation of the heterodimeric transport complex, yielding 30 mg of TAP/liter of culture. Detailed analysis of TAP function in the membrane, solubilized, purified, and reconstituted states revealed a direct influence of the native lipid environment on activity. TAP-associated phospholipids, essential for function, were profiled by liquid chromatography Fourier transform mass spectrometry. The antigen translocation activity is stimulated by phosphatidylinositol and -ethanolamine, whereas cholesterol has a negative effect on TAP activity.
KW  - ABC Transporter
KW  - Antigen Processing
KW  - ATPases
KW  - Peptides
KW  - Reconstitution of Membrane Transporters
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/76580
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-765801
SN  - 0021-9258
VL  - 286.2011
IS  - 15
SP  - 13346
EP  - 13356
PB  - American Society for Biochemistry and Molecular Biology Publications
CY  - Bethesda, Md
ER  -