TY  - JOUR
AU  - Glück, J.M.
AU  - Koenig, B. W.
AU  - Willbold, D.
TI  - Nanodiscs allow the use of integral membrane proteins as analytes in surface plasmon resonance studies
JO  - Analytical biochemistry
VL  - 408
SN  - 0003-2697
CY  - San Diego, Calif.
PB  - Elsevier
M1  - PreJuSER-11819
SP  - 46 - 52
PY  - 2011
N1  - Record converted from VDB: 12.11.2012
AB  - 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.
KW  - Antibodies, Monoclonal: immunology
KW  - Antigens, CD4: chemistry
KW  - Antigens, CD4: genetics
KW  - Antigens, CD4: metabolism
KW  - Biosensing Techniques: methods
KW  - Humans
KW  - Kinetics
KW  - Membrane Proteins: chemistry
KW  - Membrane Proteins: genetics
KW  - Membrane Proteins: metabolism
KW  - Nanostructures: chemistry
KW  - Oligopeptides: chemistry
KW  - Oligopeptides: genetics
KW  - Oligopeptides: metabolism
KW  - Protein Binding
KW  - Recombinant Fusion Proteins: chemistry
KW  - Recombinant Fusion Proteins: genetics
KW  - Recombinant Fusion Proteins: metabolism
KW  - Surface Plasmon Resonance: methods
KW  - Ubiquitin: chemistry
KW  - Ubiquitin: genetics
KW  - Ubiquitin: metabolism
KW  - Antibodies, Monoclonal (NLM Chemicals)
KW  - Antigens, CD4 (NLM Chemicals)
KW  - Membrane Proteins (NLM Chemicals)
KW  - Oligopeptides (NLM Chemicals)
KW  - Recombinant Fusion Proteins (NLM Chemicals)
KW  - Ubiquitin (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:20804721
UR  - <Go to ISI:>//WOS:000283886600007
DO  - DOI:10.1016/j.ab.2010.08.028
UR  - https://juser.fz-juelich.de/record/11819
ER  -