2LJ2

Integral membrane core domain of the mercury transporter MerF in lipid bilayer membranes

Summary for 2LJ2

• Entry DOI: 10.2210/pdb2lj2/pdb
• Related: 1WAZ 2H3O
• NMR Information: BMRB: 17914
• Descriptor: MerF (1 entity in total)
• Functional Keywords: membrane protein, mercury transporter, lipid bilayers
• Biological source: Morganella morganii
• Total number of polymer chains: 1
• Total formula weight: 6281.45

Authors

Das, B.B.,Nothnagel, H.J.,Lu, G.J.,Son, W.,Park, S.,Tian, Y.B.,Marassi, F.M.,Opella, S.J. (deposition date: 2011-09-03, release date: 2012-01-18, Last modification date: 2024-05-15)

Primary citation

Das, B.B.,Nothnagel, H.J.,Lu, G.J.,Son, W.S.,Tian, Y.,Marassi, F.M.,Opella, S.J.
Structure determination of a membrane protein in proteoliposomes.
J.Am.Chem.Soc., 134:2047-2056, 2012

PubMed Abstract: An NMR method for determining the three-dimensional structures of membrane proteins in proteoliposomes is demonstrated by determining the structure of MerFt, the 60-residue helix-loop-helix integral membrane core of the 81-residue mercury transporter MerF. The method merges elements of oriented sample (OS) solid-state NMR and magic angle spinning (MAS) solid-state NMR techniques to measure orientation restraints relative to a single external axis (the bilayer normal) from individual residues in a uniformly (13)C/(15)N labeled protein in unoriented liquid crystalline phospholipid bilayers. The method relies on the fast (>10(5) Hz) rotational diffusion of membrane proteins in bilayers to average the static chemical shift anisotropy and heteronuclear dipole-dipole coupling powder patterns to axially symmetric powder patterns with reduced frequency spans. The frequency associated with the parallel edge of such motionally averaged powder patterns is exactly the same as that measured from the single line resonance in the spectrum of a stationary sample that is macroscopically aligned parallel to the direction of the applied magnetic field. All data are collected on unoriented samples undergoing MAS. Averaging of the homonuclear (13)C/(13)C dipolar couplings, by MAS of the sample, enables the use of uniformly (13)C/(15)N labeled proteins, which provides enhanced sensitivity through direct (13)C detection as well as the use of multidimensional MAS solid-state NMR methods for resolving and assigning resonances. The unique feature of this method is the measurement of orientation restraints that enable the protein structure and orientation to be determined in unoriented proteoliposomes.

PubMed: 22217388
DOI: 10.1021/ja209464f

Experimental method

SOLID-STATE NMR