Document Detail

Expression of human ferredoxin in Saccharomyces cerevisiae: mitochondrial import of the protein and assembly of the [2Fe-2S] center.
MedLine Citation:
PMID:  1314545     Owner:  NLM     Status:  MEDLINE    
Vertebrate ferredoxins function in the transfer of reducing equivalents from NADPH:ferredoxin oxidoreductase to cytochrome P450 enzymes involved in steroid metabolism. We report here the expression of human mitochondrial ferredoxin in the yeast Saccharomyces cerevisiae. The full-length ferredoxin protein containing the ferredoxin mitochondrial leader sequence could not be stably expressed in S. cerevisiae, but a fusion protein consisting of the mature portion of ferredoxin linked to the mitochondrial leader sequence of the S. cerevisiae cytochrome c oxidase subunit Va protein (COX5a) could be stably expressed. The COX5a:ferredoxin fusion protein was targeted to the mitochondria as a preprotein and was cleaved at the normal processing site of the COX5a presequence during import into the matrix. Absorption spectra and electron transfer activity of the isolated fusion protein established that the [2Fe-2S] center was correctly assembled and incorporated into the recombinant ferredoxin in this heterologous system.
B L Seaton; L E Vickery
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Archives of biochemistry and biophysics     Volume:  294     ISSN:  0003-9861     ISO Abbreviation:  Arch. Biochem. Biophys.     Publication Date:  1992 May 
Date Detail:
Created Date:  1992-05-15     Completed Date:  1992-05-15     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0372430     Medline TA:  Arch Biochem Biophys     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  603-8     Citation Subset:  IM    
Department of Physiology and Biophysics, University of California, Irvine 92717.
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MeSH Terms
Amino Acid Sequence
Electron Transport Complex IV / biosynthesis,  genetics*,  isolation & purification
Electrophoresis, Polyacrylamide Gel
Ferredoxins / biosynthesis*,  genetics,  isolation & purification
Genes, Fungal
Mitochondria / metabolism*
Molecular Sequence Data
Molecular Weight
Promoter Regions, Genetic
Protein Sorting Signals / genetics
Recombinant Fusion Proteins / biosynthesis*,  isolation & purification
Saccharomyces cerevisiae / genetics*,  metabolism
Grant Support
Reg. No./Substance:
0/Ferredoxins; 0/Oligodeoxyribonucleotides; 0/Protein Sorting Signals; 0/Recombinant Fusion Proteins; EC Transport Complex IV

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