Reconstitution of integral membrane proteins into isotropic bicelles with improved sample stability and expanded lipid composition profile. Biochim Biophys Acta 2012 Mar;1818(3):814-20
Date
01/10/2012Pubmed ID
22226849Pubmed Central ID
PMC3292179DOI
10.1016/j.bbamem.2011.12.020Scopus ID
2-s2.0-84855774075 (requires institutional sign-in at Scopus site) 58 CitationsAbstract
Reconstitution of integral membrane proteins into membrane mimetic environments suitable for biophysical and structural studies has long been a challenge. Isotropic bicelles promise the best of both worlds-keeping a membrane protein surrounded by a small patch of bilayer-forming lipids while remaining small enough to tumble isotropically and yield good solution NMR spectra. However, traditional methods for the reconstitution of membrane proteins into isotropic bicelles expose the proteins to potentially destabilizing environments. Reconstituting the protein into liposomes and then adding short-chain lipid to this mixture produces bicelle samples while minimizing protein exposure to unfavorable environments. The result is higher yield of protein reconstituted into bicelles and improved long-term stability, homogeneity, and sample-to-sample reproducibility. This suggests better preservation of protein structure during the reconstitution procedure and leads to decreased cost per sample, production of fewer samples, and reduction of the NMR time needed to collect a high quality spectrum. Furthermore, this approach enabled reconstitution of protein into isotropic bicelles with a wider range of lipid compositions. These results are demonstrated with the small multidrug resistance transporter EmrE, a protein known to be highly sensitive to its environment.
Author List
Morrison EA, Henzler-Wildman KAAuthor
Emma A. Morrison PhD Associate Professor in the Biochemistry department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AntiportersDrug Resistance, Multiple, Bacterial
Escherichia coli
Escherichia coli Proteins
Liposomes
Membrane Lipids
Nuclear Magnetic Resonance, Biomolecular
Protein Stability
Protein Structure, Tertiary
