Document Detail

Functional analysis of a genetic defect of copper transport (Menkes disease) in different cell lines.
MedLine Citation:
PMID:  8760068     Owner:  NLM     Status:  MEDLINE    
To define the function of the Cu-transporting ATPase in Menkes disease, Menkes and normal fibroblasts were incubated with 67Cu before and after brief exposure to -SH reagents, p-chloromercuribenzoate (PCMB) and dithiothreitol (DTT). Accumulation and retention were compared among these cells, BeWo cells, and rat C6 glioma cells similarly treated. The Michaelis constant for influx of 67Cu into normal and Menkes fibroblasts was practically the same (0.21 +/- 0.07 vs. 0.24 +/- 0.06 microM). The PCMB treatment stimulated 67Cu accumulation in C6 cells, inhibited accumulation in normal and Menkes fibroblasts, and did not affect BeWo cells. DTT stimulated 67Cu uptake in all cells but BeWo cells. DTT treatment after PCMB further enhanced 67Cu accumulation in normal fibroblasts and C6 cells but had no enhancing effect on Menkes fibroblasts or BeWo cells. Menkes fibroblasts and BeWo cells released 67Cu at rates considerably slower than normal fibroblasts (0.06 and 0.09 vs. 0.22%/min, respectively). The PCMB blocked 67Cu release from normal fibroblasts but did not affect Menkes fibroblasts or BeWo cells. Reverse transcription-polymerase chain reaction analysis of total RNA from BeWo cells failed to show a predicted 943-base pair fragment representing a partial transcript of the Menkes factor. The fragment was present in extracts from normal fibroblasts. We conclude that the mechanism underlying Cu homeostasis varies among different cell types. As exemplified by BeWo and Menkes cells, failure to efflux Cu ions may be linked with the failure to express a functional Cu-transporting ATPase, namely, the Menkes protein.
Y Qian; E Tiffany-Castiglioni; E D Harris
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The American journal of physiology     Volume:  271     ISSN:  0002-9513     ISO Abbreviation:  Am. J. Physiol.     Publication Date:  1996 Jul 
Date Detail:
Created Date:  1996-12-03     Completed Date:  1996-12-03     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370511     Medline TA:  Am J Physiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  C378-84     Citation Subset:  IM    
Department of Biochemistry, Texas A&M University, College Station 77843, USA.
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MeSH Terms
Biological Transport
Cell Line
Chloromercuribenzoates / pharmacology
Copper / metabolism*
Dithiothreitol / pharmacology
Fibroblasts / drug effects,  metabolism
Menkes Kinky Hair Syndrome / metabolism*,  pathology
Polymerase Chain Reaction
Reference Values
Sulfhydryl Reagents / pharmacology
Transcription, Genetic
p-Chloromercuribenzoic Acid
Grant Support
Reg. No./Substance:
0/Chloromercuribenzoates; 0/Sulfhydryl Reagents; 3483-12-3/Dithiothreitol; 59-85-8/p-Chloromercuribenzoic Acid; 7440-50-8/Copper

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