Journal Article

PreJuSER-7145

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Integral membrane proteins in nanodiscs can be studied by solution NMR spectroscopy

Glück, J. M.FZJ* ; Wittlich, M.FZJ* ; Feuerstein, S. E.FZJ* ; Hoffmann, S.FZJ* ; Willbold, D.FZJ* ; Koenig, B. W.FZJ*

2009
American Chemical Society Washington, DC

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Please use a persistent id in citations: doi:10.1021/ja904897p

Abstract: We present a two-dimensional solution NMR spectrum of an integral membrane protein (IMP) in a nanodisc. Solution NMR relies on rapid isotropic tumbling of the analyte with correlation times in the nanosecond range. IMPs in a cellular membrane do not satisfy this condition. Previous liquid-state NMR studies on IMPs were conducted in organic solvent or artificial membrane mimicking particles like detergent micelles. Nanodiscs are relatively small (150 kDa), detergent-free model membranes that are suitable for functional reconstitution of IMPs. Nanodiscs allow solubilization of integral membrane proteins in a nearly native lipid bilayer environment. The 70 residue polypeptide CD4mut was incorporated into nanodiscs. CD4mut features one transmembrane helix. The aliphatic (1)H-(13)C HSQC spectrum of nanodiscs with inserted, ((13)C, (15)N)-labeled CD4mut exhibits reasonably dispersed protein and lipid NMR signals. Our results demonstrate that IMPs in nanodiscs are amenable to liquid-state NMR methodology.

Keyword(s): Antigens, CD4: chemistry (MeSH) ; Antigens, CD4: genetics (MeSH) ; Humans (MeSH) ; Magnetic Resonance Spectroscopy (MeSH) ; Membrane Proteins: chemistry (MeSH) ; Membrane Proteins: genetics (MeSH) ; Membranes, Artificial (MeSH) ; Nanostructures: chemistry (MeSH) ; Solutions (MeSH) ; Antigens, CD4 ; Membrane Proteins ; Membranes, Artificial ; Solutions ; J

Note: We thank Matthias Stoldt and Rudolf Hartmann for expert assistance and discussions. This work was supported by a grant from the Helmholtz Society ("Virtual Institute for Structural Biology") to D.W.

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Contributing Institute(s):

1. Strukturbiochemie (ISB-3)
2. Jülich Aachen Research Alliance - High-Performance Computing (JARA-HPC)

Research Program(s):

1. Funktion und Dysfunktion des Nervensystems (P33)

Appears in the scientific report 2009

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