Document Detail


Identification of three cysteines as targets for the Zn2+ blockade of the human skeletal muscle chloride channel.
MedLine Citation:
PMID:  10206982     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Currents through the human skeletal muscle chloride channel hClC-1 can be blocked by external application of 1 mM Zn2+ or the histidine-reactive compound diethyl pyrocarbonate (DEPC). The current block by Zn2+ strongly depends on the external pH (pKa near 6.9), whereas the block by DEPC is rather independent of the pH in the range of 5.5 to 8.5. To identify the target sites of these reagents, we constructed a total of twelve cysteine- and/or histidine-replacement mutants, transfected tsA201 cells with them, and investigated the resulting whole-cell chloride currents. The majority of the mutants exhibited a similar sensitivity toward Zn2+ or DEPC as wild type (WT) channels. Block by 1 mM Zn2+ was nearly absent only with the mutant C546A. Four mutants (C242A, C254A, H180A, and H451A) were slightly less sensitive to Zn2+ than WT. Tests with double, triple, and quadruple mutants yielded that, in addition to C546, C242 and C254 are also most likely participating in Zn2+-binding.
Authors:
L L Kürz; H Klink; I Jakob; M Kuchenbecker; S Benz; F Lehmann-Horn; R Rüdel
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  274     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1999 Apr 
Date Detail:
Created Date:  1999-05-20     Completed Date:  1999-05-20     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  11687-92     Citation Subset:  IM    
Affiliation:
Departments of General and Applied Physiology, University of Ulm, D-89069 Ulm, Germany. loathar.kuerz@medizin.uni-ulm.de
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MeSH Terms
Descriptor/Qualifier:
Cell Line
Chloride Channels / antagonists & inhibitors*,  genetics
Cysteine / genetics,  metabolism*
Diethyl Pyrocarbonate / pharmacology
Histidine / genetics,  metabolism
Humans
Hydrogen-Ion Concentration
Muscle Proteins / antagonists & inhibitors*,  genetics
Muscle, Skeletal / cytology,  drug effects*,  metabolism
Mutagenesis, Site-Directed
Zinc / pharmacology*
Chemical
Reg. No./Substance:
0/CLC-1 channel; 0/Chloride Channels; 0/Muscle Proteins; 1609-47-8/Diethyl Pyrocarbonate; 52-90-4/Cysteine; 71-00-1/Histidine; 7440-66-6/Zinc

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


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