%0 Journal Article
%A Glück, J.M.
%A Koenig, B. W.
%A Willbold, D.
%T Nanodiscs allow the use of integral membrane proteins as analytes in surface plasmon resonance studies
%J Analytical biochemistry
%V 408
%@ 0003-2697
%C San Diego, Calif.
%I Elsevier
%M PreJuSER-11819
%P 46 - 52
%D 2011
%Z Record converted from VDB: 12.11.2012
%X 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.
%K Antibodies, Monoclonal: immunology
%K Antigens, CD4: chemistry
%K Antigens, CD4: genetics
%K Antigens, CD4: metabolism
%K Biosensing Techniques: methods
%K Humans
%K Kinetics
%K Membrane Proteins: chemistry
%K Membrane Proteins: genetics
%K Membrane Proteins: metabolism
%K Nanostructures: chemistry
%K Oligopeptides: chemistry
%K Oligopeptides: genetics
%K Oligopeptides: metabolism
%K Protein Binding
%K Recombinant Fusion Proteins: chemistry
%K Recombinant Fusion Proteins: genetics
%K Recombinant Fusion Proteins: metabolism
%K Surface Plasmon Resonance: methods
%K Ubiquitin: chemistry
%K Ubiquitin: genetics
%K Ubiquitin: metabolism
%K Antibodies, Monoclonal (NLM Chemicals)
%K Antigens, CD4 (NLM Chemicals)
%K Membrane Proteins (NLM Chemicals)
%K Oligopeptides (NLM Chemicals)
%K Recombinant Fusion Proteins (NLM Chemicals)
%K Ubiquitin (NLM Chemicals)
%K J (WoSType)
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:20804721
%U <Go to ISI:>//WOS:000283886600007
%R 10.1016/j.ab.2010.08.028
%U https://juser.fz-juelich.de/record/11819