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@ARTICLE{Glck:11819,
author = {Glück, J.M. and Koenig, B. W. and Willbold, D.},
title = {{N}anodiscs allow the use of integral membrane proteins as
analytes in surface plasmon resonance studies},
journal = {Analytical biochemistry},
volume = {408},
issn = {0003-2697},
address = {San Diego, Calif.},
publisher = {Elsevier},
reportid = {PreJuSER-11819},
pages = {46 - 52},
year = {2011},
note = {Record converted from VDB: 12.11.2012},
abstract = {Nanodiscs are small-sized and flat model membranes that
provide a close to native environment for reconstitution of
integral membrane proteins. Incorporation of membrane
proteins into nanodiscs results in water-soluble proteolipid
particles making the membrane proteins amenable to a
multitude of bioanalytical techniques originally developed
for soluble proteins. The transmembrane domain of the human
CD4 receptor was fused to ubiquitin with a preceding
N-terminal decahistidine tag. The resulting integral
membrane protein was incorporated into nanodiscs. Binding of
the nanodisc-inserted histidine-tagged protein to a
monoclonal anti-pentahistidine antibody was quantified using
surface plasmon resonance (SPR) experiments. For the first
time, a membrane-inserted transmembrane protein was employed
as analyte while the antibody served as ligand immobilized
on the sensor chip surface. SPR experiments were conducted
in single-cycle mode. We demonstrate that the
nanodisc-incorporated membrane protein showed nearly
identical affinity toward the antibody as did the soluble
decahistidine-tagged ubiquitin studied in a comparative
experiment. Advantages of the new experimental setup and
potential applications are discussed.},
keywords = {Antibodies, Monoclonal: immunology / Antigens, CD4:
chemistry / Antigens, CD4: genetics / Antigens, CD4:
metabolism / Biosensing Techniques: methods / Humans /
Kinetics / Membrane Proteins: chemistry / Membrane Proteins:
genetics / Membrane Proteins: metabolism / Nanostructures:
chemistry / Oligopeptides: chemistry / Oligopeptides:
genetics / Oligopeptides: metabolism / Protein Binding /
Recombinant Fusion Proteins: chemistry / Recombinant Fusion
Proteins: genetics / Recombinant Fusion Proteins: metabolism
/ Surface Plasmon Resonance: methods / Ubiquitin: chemistry
/ Ubiquitin: genetics / Ubiquitin: metabolism / Antibodies,
Monoclonal (NLM Chemicals) / Antigens, CD4 (NLM Chemicals) /
Membrane Proteins (NLM Chemicals) / Oligopeptides (NLM
Chemicals) / Recombinant Fusion Proteins (NLM Chemicals) /
Ubiquitin (NLM Chemicals) / J (WoSType)},
cin = {ICS-6},
ddc = {570},
cid = {I:(DE-Juel1)ICS-6-20110106},
pnm = {Funktion und Dysfunktion des Nervensystems / BioSoft:
Makromolekulare Systeme und biologische
Informationsverarbeitung},
pid = {G:(DE-Juel1)FUEK409 / G:(DE-Juel1)FUEK505},
shelfmark = {Biochemical Research Methods / Biochemistry $\&$ Molecular
Biology / Chemistry, Analytical},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:20804721},
UT = {WOS:000283886600007},
doi = {10.1016/j.ab.2010.08.028},
url = {https://juser.fz-juelich.de/record/11819},
}