%0 Journal Article %1 Dorsch2009 %A Dorsch, S. %A Klotz, K. N. %A Engelhardt, S. %A Lohse, M. J. %A Bunemann, M. %D 2009 %J Nat Methods %K Activin After Animals Antibodies/immunology Antigens, Bacterial Binding/physiology CD28/genetics/metabolism CD86/genetics/metabolism Cardiac/metabolism Cell Fluorescence Fluorescence/*methods Fluorescent Fusion Green Humans II/metabolism Line Luminescent Mice Microscopy, Myocytes, Photobleaching/*methods Protein Proteins/genetics Proteins/genetics/immunology Proteins/genetics/metabolism Rats Recombinant Recovery Surface/genetics/*metabolism Transfection Type beta-1/genetics/metabolism beta-2/genetics/metabolism Receptor Adrenergic %N 3 %P 225-30 %T Analysis of receptor oligomerization by FRAP microscopy %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19234451 %V 6 %X Here we describe an approach to investigate di- or oligomerization of transmembrane receptors in living cells with fluorescence recovery after photobleaching (FRAP). We immobilized a defined fraction of receptors with antibodies and then measured lateral mobility of the nonimmobilized fraction by FRAP. We validated this approach with CD86 and CD28 as monomeric and dimeric reference proteins, respectively. Di- or oligomerization of G protein-coupled receptors is strongly debated. We studied human beta-adrenergic receptors as prototypical G protein-coupled receptors and found that beta(1)-AR shows transient interactions whereas beta(2)-AR can form stable oligomers. We propose that this FRAP method can be widely applied to study di- or oligomerization of cell-surface proteins.

@article{Dorsch2009, abstract = {Here we describe an approach to investigate di- or oligomerization of transmembrane receptors in living cells with fluorescence recovery after photobleaching (FRAP). We immobilized a defined fraction of receptors with antibodies and then measured lateral mobility of the nonimmobilized fraction by FRAP. We validated this approach with CD86 and CD28 as monomeric and dimeric reference proteins, respectively. Di- or oligomerization of G protein-coupled receptors is strongly debated. We studied human beta-adrenergic receptors as prototypical G protein-coupled receptors and found that beta(1)-AR shows transient interactions whereas beta(2)-AR can form stable oligomers. We propose that this FRAP method can be widely applied to study di- or oligomerization of cell-surface proteins.}, added-at = {2010-12-14T18:12:02.000+0100}, author = {Dorsch, S. and Klotz, K. N. and Engelhardt, S. and Lohse, M. J. and Bunemann, M.}, biburl = {https://www.bibsonomy.org/bibtex/22ceb1103f777e6c336ab2b5c94f64d88/pharmawuerz}, endnotereftype = {Journal Article}, interhash = {a006b7664e83653b44236427cea27729}, intrahash = {2ceb1103f777e6c336ab2b5c94f64d88}, issn = {1548-7105 (Electronic) 1548-7105 (Linking)}, journal = {Nat Methods}, keywords = {Activin After Animals Antibodies/immunology Antigens, Bacterial Binding/physiology CD28/genetics/metabolism CD86/genetics/metabolism Cardiac/metabolism Cell Fluorescence Fluorescence/*methods Fluorescent Fusion Green Humans II/metabolism Line Luminescent Mice Microscopy, Myocytes, Photobleaching/*methods Protein Proteins/genetics Proteins/genetics/immunology Proteins/genetics/metabolism Rats Recombinant Recovery Surface/genetics/*metabolism Transfection Type beta-1/genetics/metabolism beta-2/genetics/metabolism Receptor Adrenergic}, month = Mar, note = {Dorsch, Sandra Klotz, Karl-Norbert Engelhardt, Stefan Lohse, Martin J Bunemann, Moritz Research Support, Non-U.S. Gov't United States Nature methods Nat Methods. 2009 Mar;6(3):225-30. Epub 2009 Feb 22.}, number = 3, pages = {225-30}, shorttitle = {Analysis of receptor oligomerization by FRAP microscopy}, timestamp = {2010-12-14T18:22:39.000+0100}, title = {Analysis of receptor oligomerization by FRAP microscopy}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=19234451}, volume = 6, year = 2009 }