Site-specific, orthogonal labeling of proteins in intact cells with
two small biarsenical fluorophores
A. Zurn, C. Klenk, U. Zabel, S. Reiner, M. Lohse, und C. Hoffmann. Bioconjug Chem, 21 (5):
853-9(Mai 2010)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..
Zusammenfassung
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.
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.
%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
}