Photoaffinity Labeling in Biological Signal Transduction

Dormán, György

doi:10.1007/3-540-45035-1_6

Cited by 31 publications

(12 citation statements)

References 125 publications

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“…The labeled site can then be identified using any of a number of biochemical techniques, for instance limited proteolysis followed by gel electrophoresis and/or mass spectrometry to suggest the location of the binding site for the unlabeled ligand. ( 14 )…”

Section: Resultsmentioning

confidence: 99%

“…Under UV irradiation, aryl azides extrude a molecule of N 2 , transiently forming a reactive nitrene intermediate, which rearranges to an electrophilic cyclic ketenimine. ( 14 ) In photoaffinity labeling experiments, this occurs within a protein binding site, and the ketenimine is quenched by a nucleophilic functional group from the protein, thus forming a covalent linkage between the label and the protein (Scheme 2 a). To demonstrate that azide 11 is capable of undergoing this chemistry, a benzene solution of 11 , containing an excess of diethylamine, was irradiated with an unfiltered (full spectrum) mercury lamp (Scheme 2 b).…”

Section: Resultsmentioning

confidence: 99%

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Synthesis and Properties of Molecular Probes for the Rescue Site on Mutant Cystic Fibrosis Transmembrane Conductance Regulator

et al. 2011

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Cystic fibrosis is a genetic disease caused by mutations in the gene for the cystic fibrosis transmembrane conductance regulator (CFTR) protein. In vitro experiments have demonstrated that 4-methyl-2-(5-phenyl-1H-pyrazol-3-yl)phenol (VRT-532, 1) is able to partially restore the function of mutant CFTR proteins. To help elucidate the nature of the interactions between 1 and mutant CFTR, molecular probes based on the structure of 1 have been prepared. These include a photoreactive aryl azide derivative 11 and a fluorescent dansyl sulfonamide 15. Additionally, a method for hydrogen isotope exchange on 1 has been developed, which could be used for the incorporation of radioactive tritium. Using iodide efflux assays, the probe molecules have been demonstrated to modulate the activity of mutant CFTR in the same manner as 1. These probe molecules enable a number of biochemical experiments aimed at understanding how 1 rescues the function of mutant CFTR. This understanding can in turn aid in the design and development of more efficacious compounds which may serve as therapeutic agents in the treatment of cystic fibrosis.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Synthesis and Properties of Molecular Probes for the Rescue Site on Mutant Cystic Fibrosis Transmembrane Conductance Regulator

et al. 2011

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“…A brief comparison of benefits and downsides of commonly used photoreactive groups is presented in Table . For more detailed descriptions of these photoreactive groups and their use in PAL, see Brunner (), Dormán and Prestwich (), Dormán (), and Sadakane and Hatanaka ().…”

Section: Strategic Planningmentioning

confidence: 99%

Target Identification by Diazirine Photo‐Cross‐Linking and Click Chemistry

MacKinnon

Taunton

2009

CP Chemical Biology

118

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Target identification is often the rate‐determining step in deciphering the mechanism of action of biologically active small molecules. Photo‐affinity labeling (PAL) represents a useful biochemical strategy for target identification in complex protein mixtures. This unit describes the use of alkyl diazirine‐based photo‐affinity probes and Cu(I)‐catalyzed click chemistry to covalently label and visualize the targets of biologically active small molecules. A general method for affinity purification of probe‐modified proteins, useful for identification of protein targets, is also described. Curr. Protoc. Chem Biol. 1:55‐73. © 2009 by John Wiley & Sons, Inc.

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“…All members of this chemical compound library carry photoreactive groups. Those compounds, which are able to bind to the target protein, either form a new covalent linkage (photoaffinity labeling and photoimmobilization) or cleaving a specific bond (photodeprotection) initiated by irradiation of the photophore at a defined wavelength (DORMAN, 2000).…”

Section: Covalent Marker Librariesmentioning

confidence: 99%