title: Antigenic Peptide Recognition on the Human ABC Transporter TAP Resolved by DNP-Enhanced Solid-State NMR Spectroscopy.
abstract: The human transporter associated with antigen processing (TAP) is a 150 kDa heterodimeric ABC transport complex that selects peptides for export into the endoplasmic reticulum and subsequent loading onto major histocompatibility complex class I molecules to trigger adaptive immune responses against virally or malignantly transformed cells. To date, no atomic-resolution information on peptide-TAP interactions has been obtained, hampering a mechanistic understanding of the early steps of substrate translocation catalyzed by TAP. Here, we developed a mild method to concentrate an unstable membrane protein complex and combined this effort with dynamic nuclear polarization enhanced magic angle spinning solid-state NMR to study this challenging membrane protein-substrate complex. We were able to determine the atomic-resolution backbone conformation of an antigenic peptide bound to human TAP. Our NMR data also provide unparalleled insights into the nature of the interactions between the side chains of the antigen peptide and TAP. By combining NMR data and molecular modeling, the location of the peptide binding cavity has been identified, revealing a complex scenario of peptide-TAP recognition. Our findings reveal a structural and chemical basis of substrate selection rules, which define the crucial function of this ABC transporter in human immunity and health. This work is the first NMR study of a eukaryotic transporter protein and presents the power of solid-state NMR in this growing field.
authors.forename: Elisa
authors.initials: E
authors.lastname: Lehnert
authors.forename: Jiafei
authors.initials: J
authors.lastname: Mao
authors.affiliation: Department of Theoretical Biophysics, Max Planck Institute of Biophysics , Max-von-Laue-Str. 3, 60438 Frankfurt am Main, Germany.
authors.forename: Ahmad Reza
authors.initials: AR
authors.lastname: Mehdipour
authors.affiliation: Department of Theoretical Biophysics, Max Planck Institute of Biophysics , Max-von-Laue-Str. 3, 60438 Frankfurt am Main, Germany.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Hummer
authors.forename: Rupert
authors.initials: R
authors.lastname: Abele
authors.forename: Clemens
authors.initials: C
authors.lastname: Glaubitz
authors.forename: Robert
authors.initials: R
authors.lastname: Tampé