title: Modified lipid and protein dynamics in nanodiscs.
abstract: For membrane protein studies, nanodiscs have been shown to hold great potential in terms of preparing soluble samples while maintaining a lipid environment. Here, we describe the differences in lipid order and protein dynamics in MSP1 nanodiscs compared to lamellar preparations by solid-state NMR. For DMPC, an increase of the dipolar C-H lipid acyl chain order parameters in nanodiscs is observed in both gel- and liquid crystalline phases. Incorporating proteorhodopsin in these nanodiscs resulted in a significantly longer rotating frame spin-lattice relaxation time for (13)C leerzeichen and better cross polarisation efficiency due to restricted protein dynamics. A comparison of (13)C-(13)C correlation spectra revealed no structural differences. The incorporation of proteorhodopsin into nanodiscs has been optimised with respect to detergent and to protein/scaffold protein/lipid stoichiometries. Its functional state was probed by time-resolved optical spectroscopy revealing only minor differences between lamellar and nanodisc preparations. Our observations show remarkable dynamic effects between membrane proteins, lipids and scaffold protein. The potential use of nanodiscs for solid-state NMR applications is discussed.
authors.affiliation: Institute of Biophysical Chemistry and Centre for Biomolecular Magnetic Resonance, Goethe University Frankfurt, 60438 Frankfurt/M, Germany.
authors.forename: Karsten
authors.initials: K
authors.lastname: Mörs
authors.forename: Christian
authors.initials: C
authors.lastname: Roos
authors.forename: Frank
authors.initials: F
authors.lastname: Scholz
authors.forename: Josef
authors.initials: J
authors.lastname: Wachtveitl
authors.forename: Volker
authors.initials: V
authors.lastname: Dötsch
authors.forename: Frank
authors.initials: F
authors.lastname: Bernhard
authors.forename: Clemens
authors.initials: C
authors.lastname: Glaubitz
mesh_terms: D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008563:Membrane Lipids / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D012243:Rhodopsin / Q000737:chemistry; D025604:Rhodopsins, Microbial
chemical_list: D008563:Membrane Lipids; D008565:Membrane Proteins; D025604:Rhodopsins, Microbial; C413548:proteorhodopsin; D012243:Rhodopsin; D004134:Dimyristoylphosphatidylcholine