Membrane proteins solubilized intact in lipid containing nanoparticles bounded by styrene maleic acid copolymer.

One-third of eukaryotic proteins are integrated within membranes, as are the targets of 40% of approved drugs. However, the lack of a general means of solubilizing, stabilizing and structurally characterizing active membrane proteins has frustrated efforts to understand their mechanisms and exploit their potential value. Here we report that bilayer disks formed by phospholipids and styrene maleic anhydride copolymer preserve the functional and structural integrity of alpha-helical and beta-barrel transmembrane proteins. They form 11 nm particles that are monodispersed, biocompatible, thermostable, and water-soluble, allowing diverse membrane proteins to be simply and rapidly presented for virtually any in vitro analysis.

[1]  E. Beckmann,et al.  Lipid location in deoxycholate-treated purple membrane at 2.6 A. , 1995, Journal of molecular biology.

[2]  A. Plückthun,et al.  Acyl and phosphoryl migration in lysophospholipids: importance in phospholipid synthesis and phospholipase specificity. , 1982, Biochemistry.

[3]  Wing-Yiu Choy,et al.  Solution structure and dynamics of the outer membrane enzyme PagP by NMR , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[4]  B. Wallace,et al.  Circular dichroism analyses of membrane proteins: an examination of differential light scattering and absorption flattening effects in large membrane vesicles and membrane sheets. , 1984, Analytical biochemistry.

[5]  Stephen G. Sligar,et al.  Self-Assembly of Discoidal Phospholipid Bilayer Nanoparticles with Membrane Scaffold Proteins , 2002 .

[6]  C. Sanders,et al.  Amphipols can support the activity of a membrane enzyme. , 2002, Journal of the American Chemical Society.

[7]  S. Sligar,et al.  Self‐assembly of single integral membrane proteins into soluble nanoscale phospholipid bilayers , 2003, Protein science : a publication of the Protein Society.

[8]  L. Kay,et al.  The integral membrane enzyme PagP alternates between two dynamically distinct states. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[9]  Samuel I. Miller,et al.  Transfer of palmitate from phospholipids to lipid A in outer membranes of Gram‐negative bacteria , 2000, The EMBO journal.

[10]  Samuel I. Miller,et al.  Lipid A Acylation and Bacterial Resistance against Vertebrate Antimicrobial Peptides , 1998, Cell.

[11]  B. Tighe,et al.  Responsive hydrophobically associating polymers: a review of structure and properties. , 2001, Advanced drug delivery reviews.

[12]  G. Privé,et al.  Lipopeptide detergents designed for the structural study of membrane proteins , 2003, Nature Biotechnology.

[13]  P. Bauer,et al.  A natural CD label to probe the structure of the purple membrane from Halobacterium halobium by means of exciton coupling effects. , 1975, Biochemical and biophysical research communications.