This title appears in the Scientific Report :
2011
Please use the identifier:
http://dx.doi.org/10.1016/j.ab.2010.08.028 in citations.
Nanodiscs allow the use of integral membrane proteins as analytes in surface plasmon resonance studies
Nanodiscs allow the use of integral membrane proteins as analytes in surface plasmon resonance studies
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 bio...
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Personal Name(s): | Glück, J.M. |
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Koenig, B. W. / Willbold, D. | |
Contributing Institute: |
Strukturbiochemie; ICS-6 |
Published in: | Analytical biochemistry, 408 (2011) S. 46 - 52 |
Imprint: |
San Diego, Calif.
Elsevier
2011
|
Physical Description: |
46 - 52 |
DOI: |
10.1016/j.ab.2010.08.028 |
PubMed ID: |
20804721 |
Document Type: |
Journal Article |
Research Program: |
BioSoft: Makromolekulare Systeme und biologische Informationsverarbeitung Funktion und Dysfunktion des Nervensystems |
Series Title: |
Analytical Biochemistry
408 |
Subject (ZB): | |
Publikationsportal JuSER |
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. |