NMR solution structure determination of membrane proteins reconstituted in detergent micelles.
As an alternative to X-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy in solution can be used for three-dimensional structure determination of small membrane proteins, preferably proteins with beta-barrel fold. This paper reviews recent achievements as well as limiting factors encountered in solution NMR studies of membrane proteins. Our particular interest has been focused on supplementing structure determination with data on the solvation of the proteins in the mixed micelles with detergents that are used to reconstitute membrane proteins for the NMR experiments. For the Escherichia coli outer membrane protein X (OmpX) in dihexanoylphosphatidylcholine (DHPC) micelles, such studies showed that the central part of the protein is covered with a fluid monolayer of lipid molecules, which seems to mimic quite faithfully the embedding of the protein in the lipid phase of the biological membrane. The implication is that the micellar systems used in this instance for the NMR studies of the membrane protein should also be suitable for further investigations of functional interactions with other proteins or low-molecular weight ligands.
Fernández
César
C
Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule Zürich, CH-8093 Zürich, Switzerland. cesar.fernandez@pharma.novartis.com
Wüthrich
Kurt
K
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry; D003902:Detergents; D029968:Escherichia coli Proteins / Q000737:chemistry; D006867:Hydrolases; D008565:Membrane Proteins / Q000737:chemistry*; D008823:Micelles; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D010742:Phospholipid Ethers; D011487:Protein Conformation; D012996:Solutions
C052298:1,2-dihexadecyl-sn-glycero-3-phosphocholine; D001425:Bacterial Outer Membrane Proteins; D003902:Detergents; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D008823:Micelles; D010742:Phospholipid Ethers; D012996:Solutions; C482905:OmpX protein, E coli; D006867:Hydrolases