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Title: “Turn-On” Protein Fluorescence: In Situ Formation of Cyanine Dyes

Journal Article · · Journal of the American Chemical Society
DOI: https://doi.org/10.1021/ja506376j · OSTI ID:1237768

Protein reengineering of cellular retinoic acid binding protein II (CRABPII) has yielded a genetically addressable system, capable of binding a profluorophoric chromophore that results in fluorescent protein/chromophore complexes. These complexes exhibit far-red emission, with high quantum efficiencies and brightness and also exhibit excellent pH stability spanning the range of 2$$-$$11. In the course of this study, it became evident that single mutations of L121E and R59W were most effective in improving the fluorescent characteristics of CRABPII mutants as well as the kinetics of complex formation. The readily crystallizable nature of these proteins was invaluable to provide clues for the observed spectroscopic behavior that results from single mutation of key residues.

Research Organization:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Grant/Contract Number:
W-31109-ENG-38
OSTI ID:
1237768
Journal Information:
Journal of the American Chemical Society, Journal Name: Journal of the American Chemical Society Journal Issue: 3 Vol. 137; ISSN 0002-7863
Publisher:
American Chemical Society (ACS)Copyright Statement
Country of Publication:
United States
Language:
ENGLISH

References (81)

Analysis of a data set of paired uncomplexed protein structures: New metrics for side-chain flexibility and model evaluation journal January 2001
Ultrafast Radiationless Deactivation of Organic Dyes:  Evidence for a Two-State Two-Mode Pathway in Polymethine Cyanines journal March 2000
Bioorthogonal Reactions for Labeling Proteins journal October 2013
Structures of cellular retinoic acid binding proteins I and II in complex with synthetic retinoids journal November 1999
An improved monomeric infrared fluorescent protein for neuronal and tumour brain imaging journal May 2014
Fluorogen-activating proteins as biosensors of cell-surface proteins in living cells journal June 2010
Chemical Tags for Labeling Proteins Inside Living Cells journal September 2011
Bioorthogonal Chemistry: Fishing for Selectivity in a Sea of Functionality journal September 2009
An Engineered Protein Tag for Multiprotein Labeling in Living Cells journal February 2008
A guide to choosing fluorescent proteins journal November 2005
Rational Design of a Colorimetric pH Sensor from a Soluble Retinoic Acid Chaperone journal October 2013
Areas, Volumes, Packing, and Protein Structure journal June 1977
Development of SNAP-Tag Fluorogenic Probes for Wash-Free Fluorescence Imaging journal July 2011
Specific Covalent Labeling of Recombinant Protein Molecules Inside Live Cells journal July 1998
Photophysics of Cy3-Encapsulated Calcium Phosphate Nanoparticles journal April 2009
High-throughput flow cytometry compatible biosensor based on fluorogen activating protein technology journal January 2013
Genetically Encoded Cyclopropene Directs Rapid, Photoclick-Chemistry-Mediated Protein Labeling in Mammalian Cells journal September 2012
Mutational Analysis of Deinococcus radiodurans Bacteriophytochrome Reveals Key Amino Acids Necessary for the Photochromicity and Proton Exchange Cycle of Phytochromes journal January 2008
A Rapid SNAP-Tag Fluorogenic Probe Based on an Environment-Sensitive Fluorophore for No-Wash Live Cell Imaging journal August 2014
Fluorescent Protein Tracking and Detection: Fluorescent Protein Structure and Color Variants journal December 2009
Fluorescence Visualization of Newly Synthesized Proteins in Mammalian Cells journal November 2006
Amino Acids for Diels-Alder Reactions in Living Cells journal March 2012
Improved green fluorescence journal February 1995
A Fluorogenic TMP-Tag for High Signal-to-Background Intracellular Live Cell Imaging journal June 2013
Computational design of a red fluorophore ligase for site-specific protein labeling in living cells journal October 2014
Tuning the Electronic Absorption of Protein-Embedded All-trans-Retinal journal December 2012
Side-chain flexibility in proteins upon ligand binding journal May 2000
Green fluorescent protein as a marker for gene expression journal February 1994
An In Vivo Covalent TMP-Tag Based on Proximity-Induced Reactivity journal July 2009
Bright and stable near-infrared fluorescent protein for in vivo imaging journal July 2011
A Strain-Promoted [3 + 2] Azide−Alkyne Cycloaddition for Covalent Modification of Biomolecules in Living Systems journal November 2004
Bioconjugation by Copper(I)-Catalyzed Azide-Alkyne [3 + 2] Cycloaddition journal March 2003
Mammalian Expression of Infrared Fluorescent Proteins Engineered from a Bacterial Phytochrome journal May 2009
Choosing the Sequence of dsRNA for RNAi in Mouse journal January 2009
Primary structure of the Aequorea victoria green-fluorescent protein journal February 1992
Structure/Function of Cytoplasmic Vitamin A-Binding Proteins journal July 1996
Diels–Alder Cycloaddition for Fluorophore Targeting to Specific Proteins inside Living Cells journal December 2011
Near-infrared fluorescent proteins for multicolor in vivo imaging journal June 2013
Copper-free click chemistry for dynamic in vivo imaging journal October 2007
Probing Wavelength Regulation with an Engineered Rhodopsin Mimic and a C15-Retinal Analogue journal February 2012
Tetrazine-Based Cycloadditions: Application to Pretargeted Live Cell Imaging journal December 2008
Chemical labeling strategies for cell biology journal July 2006
Engineering a Rhodopsin Protein Mimic journal April 2006
Experimental and Computational Investigation of Unsymmetrical Cyanine Dyes:  Understanding Torsionally Responsive Fluorogenic Dyes journal May 2007
The nature of the different environmental sensitivity of symmetrical and unsymmetrical cyanine dyes: an experimental and theoretical study journal January 2012
Polarity as a criterion in protein design journal January 1989
Fluorescent proteins at a glance journal December 2010
Photophysics of Indocarbocyanine Dyes in Organic Solvents journal April 1998
New Biarsenical Ligands and Tetracysteine Motifs for Protein Labeling in Vitro and in Vivo:  Synthesis and Biological Applications journal May 2002
HaloTag: A Novel Protein Labeling Technology for Cell Imaging and Protein Analysis journal June 2008
Polymethine dyes as spectral-fluorescent probes for biomacromolecules journal March 2012
Extraction, Purification and Properties of Aequorin, a Bioluminescent Protein from the Luminous Hydromedusan,Aequorea journal June 1962
Bioorthogonal Turn-On Probes for Imaging Small Molecules inside Living Cells journal March 2010
Chemical biology-based approaches on fluorescent labeling of proteins in live cells journal January 2013
NIR dyes for bioimaging applications journal February 2010
Far-red fluorescent tags for protein imaging in living tissues journal February 2009
Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein journal March 2014
Reversible Synthesis and Characterization of Dynamic Imino Analogues of Trimethine and Pentamethine Cyanine Dyes journal February 2009
Enzymatic Labeling of Proteins: Techniques and Approaches journal August 2013
A Rainbow of Fluoromodules: A Promiscuous scFv Protein Binds to and Activates a Diverse Set of Fluorogenic Cyanine Dyes journal September 2008
Advances in fluorescence labeling strategies for dynamic cellular imaging journal June 2014
Protein Design:  Reengineering Cellular Retinoic Acid Binding Protein II into a Rhodopsin Protein Mimic journal May 2007
A Bilirubin-Inducible Fluorescent Protein from Eel Muscle journal June 2013
A fluorophore ligase for site-specific protein labeling inside living cells journal June 2010
Fluorescence efficiency of four infrared polymethine dyes journal May 2003
Site specific protein labeling by enzymatic posttranslational modification journal January 2009
Merocyanines as Extremely Bathochromically Absorbing Chromophores in the Halobacterial Membrane Protein Bacteriorhodopsin journal August 1997
Cyanine dye labeling reagents: Sulfoindocyanine succinimidyl esters journal March 1993
A Red Cy3-Based Biarsenical Fluorescent Probe Targeted to a Complementary Binding Peptide journal July 2007
Labeling of fusion proteins with synthetic fluorophores in live cells journal June 2004
A general method for the covalent labeling of fusion proteins with small molecules in vivo journal December 2002
Selective Dye-Labeling of Newly Synthesized Proteins in Bacterial Cells journal October 2005
Elucidating the exact role of engineered CRABPII residues for the formation of a retinal protonated Schiff base journal December 2009
Development of Protein-Labeling Probes with a Redesigned Fluorogenic Switch Based on Intramolecular Association for No-Wash Live-Cell Imaging journal April 2012
Fluorogen-activating single-chain antibodies for imaging cell surface proteins journal December 2007
Cyanine dyes in biophysical research: the photophysics of polymethine fluorescent dyes in biomolecular environments journal November 2010
Development of Fluorogenic Probes for Quick No-Wash Live-Cell Imaging of Intracellular Proteins journal August 2013
Second-Generation Covalent TMP-Tag for Live Cell Imaging journal August 2012
How to obtain labeled proteins and what to do with them journal December 2010
A FRET-Based Fluorogenic Phosphine for Live-Cell Imaging with the Staudinger Ligation journal March 2008
The photochemical determinants of color vision: Revealing how opsins tune their chromophore's absorption wavelength journal October 2013

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