%0 Journal Article %1 Zurn2010 %A Zurn, A. %A Klenk, C. %A Zabel, U. %A Reiner, S. %A Lohse, M. J. %A Hoffmann, C. %D 2010 %J Bioconjug Chem %K 1/analysis/chemistry/metabolism Acid Amino Arrestins/analysis/chemistry/metabolism Binding Cell Data Dyes/*chemistry/metabolism Energy Fluorescence Fluorescent Hormone, Humans Line Molecular Parathyroid Protein Proteins/*analysis/chemistry/*metabolism Resonance Sequence Transfer/*methods Type Receptor %N 5 %P 853-9 %T Site-specific, orthogonal labeling of proteins in intact cells with two small biarsenical fluorophores %U http://www.ncbi.nlm.nih.gov/pubmed/20429545 %V 21 %X The fusion of fluorescent proteins to proteins of interest has greatly advanced fluorescence microscopy, but is often limited by their large size. Here, we report site-specific, orthogonal labeling of two cellular proteins in intact cells with two small fluorescent dyes: fluorescein arsenical hairpin binder, FlAsH, and its red analogue, ReAsH, which bind to tetracysteine motifs. Development of a sequential labeling method to two different motifs, CCPGCC and FLNCCPGCCMEP, allowed site-specific labeling with FlAsH and ReAsH, respectively. Using the cell surface receptor for parathyroid hormone and its cytosolic binding protein, beta-arrestin2, we show their selective visualization in intact cells and analyze their interaction by colocalization and fluorescence resonance energy transfer (FRET). We propose that this method may be widely applied to label intracellular proteins and to study their interactions in intact cells with minimal disturbance of their function.

@article{Zurn2010, abstract = {The fusion of fluorescent proteins to proteins of interest has greatly advanced fluorescence microscopy, but is often limited by their large size. Here, we report site-specific, orthogonal labeling of two cellular proteins in intact cells with two small fluorescent dyes: fluorescein arsenical hairpin binder, FlAsH, and its red analogue, ReAsH, which bind to tetracysteine motifs. Development of a sequential labeling method to two different motifs, CCPGCC and FLNCCPGCCMEP, allowed site-specific labeling with FlAsH and ReAsH, respectively. Using the cell surface receptor for parathyroid hormone and its cytosolic binding protein, beta-arrestin2, we show their selective visualization in intact cells and analyze their interaction by colocalization and fluorescence resonance energy transfer (FRET). We propose that this method may be widely applied to label intracellular proteins and to study their interactions in intact cells with minimal disturbance of their function.}, added-at = {2010-12-14T18:12:02.000+0100}, author = {Zurn, A. and Klenk, C. and Zabel, U. and Reiner, S. and Lohse, M. J. and Hoffmann, C.}, biburl = {https://www.bibsonomy.org/bibtex/2a71f7ad040379bb95e86331f4d4076ca/pharmawuerz}, endnotereftype = {Journal Article}, groups = {private}, interhash = {e2711ff0b74159ab9b1149659eb89fb8}, intrahash = {a71f7ad040379bb95e86331f4d4076ca}, issn = {1520-4812 (Electronic) 1043-1802 (Linking)}, journal = {Bioconjug Chem}, keywords = {1/analysis/chemistry/metabolism Acid Amino Arrestins/analysis/chemistry/metabolism Binding Cell Data Dyes/*chemistry/metabolism Energy Fluorescence Fluorescent Hormone, Humans Line Molecular Parathyroid Protein Proteins/*analysis/chemistry/*metabolism Resonance Sequence Transfer/*methods Type Receptor}, month = {May 19}, note = {Zurn, Alexander Klenk, Christoph Zabel, Ulrike Reiner, Susanne Lohse, Martin J Hoffmann, Carsten Research Support, Non-U.S. Gov't United States Bioconjugate chemistry Bioconjug Chem. 2010 May 19;21(5):853-9.}, number = 5, pages = {853-9}, shorttitle = {Site-specific, orthogonal labeling of proteins in intact cells with two small biarsenical fluorophores}, timestamp = {2010-12-14T18:20:23.000+0100}, title = {Site-specific, orthogonal labeling of proteins in intact cells with two small biarsenical fluorophores}, url = {http://www.ncbi.nlm.nih.gov/pubmed/20429545}, volume = 21, year = 2010 }