Document Detail


Dynamic copper(I) imaging in mammalian cells with a genetically encoded fluorescent copper(I) sensor.
MedLine Citation:
PMID:  20131768     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Copper, a key cofactor for many life processes, is toxic at elevated levels, and its availability is strictly controlled inside cells. Therefore, it is a challenge to visualize copper availability in the tight copper-binding environment of the cell. We report a genetically encoded fluorescent copper(I) sensor based on the copper(I)-binding-induced conformational change of a copper-responsive transcriptional regulator, Amt1. The resulting reporter, Amt1-FRET, is ratiometric, highly sensitive (K(d) = 2.5 x 10(-18) M), and selective toward copper(I). Its measured high affinity to copper(I) confirms the extremely low copper availability in yeast since Amt1 senses the upper limit of cellular copper levels in yeast and activates copper detoxification genes. Amt1-FRET operates in the dynamic range of the cellular copper buffer in mammalian cells and can report dynamic fluctuations of the cellular copper availability within minutes of perturbation. Thus, Amt1-FRET visualizes the tightly controlled copper availability in mammalian cells.
Authors:
Seraphine V Wegner; Hasan Arslan; Murat Sunbul; Jun Yin; Chuan He
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Journal of the American Chemical Society     Volume:  132     ISSN:  1520-5126     ISO Abbreviation:  J. Am. Chem. Soc.     Publication Date:  2010 Mar 
Date Detail:
Created Date:  2010-02-24     Completed Date:  2010-06-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7503056     Medline TA:  J Am Chem Soc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2567-9     Citation Subset:  IM    
Affiliation:
Department of Chemistry and Institute for Biophysical Dynamics, The University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
CHO Cells
Copper / analysis*,  metabolism
Cricetinae
Cricetulus
DNA-Binding Proteins / genetics*,  metabolism*
Escherichia coli / genetics
Fluorescence Resonance Energy Transfer / methods*
Fungal Proteins / genetics*,  metabolism*
Protein Binding
Saccharomyces cerevisiae / genetics,  metabolism*
Transcription Factors / genetics*,  metabolism*
Chemical
Reg. No./Substance:
0/DNA-Binding Proteins; 0/Fungal Proteins; 0/Transcription Factors; 141490-28-0/AMT1 protein, Candida glabrate; 7440-50-8/Copper

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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