caa a22 4500 445325526 CHVBK 20180317142719.0 cr unu---uuuuu 170323e20110901xx s 000 0 eng 10.1007/s10858-011-9551-z doi (NATIONALLICENCE)springer-10.1007/s10858-011-9551-z Topology and immersion depth of an integral membrane protein by paramagnetic rates from dissolved oxygen [Elektronische Daten] [M. Al-Abdul-Wahid, Raffaello Verardi, Gianluigi Veglia, R. Prosser] In studies of membrane proteins, knowledge of protein topology can provide useful insight into both structure and function. In this work, we present a solution NMR method for the measurement the tilt angle and average immersion depth of alpha helices in membrane proteins, from analysis of the paramagnetic relaxation rate enhancements arising from dissolved oxygen. No modification to the micelle or protein is necessary, and the topology of both transmembrane and amphipathic helices are readily determined. We apply this method to the measure the topology of a monomeric mutant of phospholamban (AFA-PLN), a 52-residue membrane protein containing both an amphipathic and a transmembrane alpha helix. In dodecylphosphocholine micelles, the amphipathic helix of AFA-PLN was found to have a tilt angle of 87°±1° and an average immersion depth of 13.2Å. The transmembrane helix was found to have an average immersion depth of 5.4Å, indicating residues 41 and 42 are closest to the micelle centre. The resolution of paramagnetic relaxation rate enhancements from dissolved oxygen compares favourably to those from Ni (II), a hydrophilic paramagnetic species. Springer Science+Business Media B.V., 2011 Membrane protein immersion depth nationallicence Membrane protein topology nationallicence Paramagnetic relaxation rate enhancements nationallicence Solution state NMR of membrane proteins nationallicence AFA-PLN : Phospholamban mutant C36A/C41F/C46A nationallicence DOPC : 1,2-dioleoyl-glycero-3-sn-phosphocholine nationallicence DOPE : Dioleylphosphoethanolamine nationallicence EPR : Electron paramagnetic resonance nationallicence HSQC : Heteronuclear single quantum correlation nationallicence NOE : Nuclear overhauser effect nationallicence PISEMA : Polarization inversion spin exchange at magic angle nationallicence PLN, wt-PLN : Phospholamban nationallicence PRE : Paramagnetic relaxation enhancement nationallicence RDC : Residual dipolar coupling nationallicence SDS : Sodium dodecyl sulfate nationallicence SERCA : Sarco(endo)plasmic reticulum calcium ATPase nationallicence Al-Abdul-Wahid M. Department of Chemistry, University of Toronto Mississauga, L5L 1C6, Mississauga, ON, Canada aut Verardi Raffaello Department of Chemistry, University of Minnesota, 55455, Minneapolis, MN, USA aut Veglia Gianluigi Department of Chemistry, University of Minnesota, 55455, Minneapolis, MN, USA aut Prosser R. Department of Chemistry, University of Toronto Mississauga, L5L 1C6, Mississauga, ON, Canada aut Journal of Biomolecular NMR Springer Netherlands 51/1-2(2011-09-01), 173-183 0925-2738 51:1-2<173 2011 51 10858 https://doi.org/10.1007/s10858-011-9551-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9551-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Al-Abdul-Wahid M. Department of Chemistry, University of Toronto Mississauga, L5L 1C6, Mississauga, ON, Canada aut NATIONALLICENCE 700 1- Verardi Raffaello Department of Chemistry, University of Minnesota, 55455, Minneapolis, MN, USA aut NATIONALLICENCE 700 1- Veglia Gianluigi Department of Chemistry, University of Minnesota, 55455, Minneapolis, MN, USA aut NATIONALLICENCE 700 1- Prosser R. Department of Chemistry, University of Toronto Mississauga, L5L 1C6, Mississauga, ON, Canada aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/1-2(2011-09-01), 173-183 0925-2738 51:1-2<173 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer