TY  - JOUR
AU  - Glück, J.M.
AU  - Wittlich, M.
AU  - Feuerstein, S.E.
AU  - Hoffmann, S.
AU  - Willbold, D.
AU  - Koenig, B. W.
TI  - Integral membrane proteins in nanodiscs can be studied by solution NMR spectroscopy
JO  - Journal of the American Chemical Society
VL  - 131
SN  - 0002-7863
CY  - Washington, DC
PB  - American Chemical Society
M1  - PreJuSER-7145
SP  - 12060 - 12061
PY  - 2009
N1  - 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.
AB  - 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.
KW  - Antigens, CD4: chemistry
KW  - Antigens, CD4: genetics
KW  - Humans
KW  - Magnetic Resonance Spectroscopy
KW  - Membrane Proteins: chemistry
KW  - Membrane Proteins: genetics
KW  - Membranes, Artificial
KW  - Nanostructures: chemistry
KW  - Solutions
KW  - Antigens, CD4 (NLM Chemicals)
KW  - Membrane Proteins (NLM Chemicals)
KW  - Membranes, Artificial (NLM Chemicals)
KW  - Solutions (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:19663495
UR  - <Go to ISI:>//WOS:000269379600020
DO  - DOI:10.1021/ja904897p
UR  - https://juser.fz-juelich.de/record/7145
ER  -