Document Detail


Recombinant expression, biophysical characterization, and cardiolipin-induced changes of two Caenorhabditis elegans cytochrome c proteins.
MedLine Citation:
PMID:  23282202     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cytochrome c (cyt c) is one of the most widely studied biomolecules, but not much is known about this protein from nematodes. Recombinant expression of Caenorhabditis elegans CYC-2.1 and CYC-2.2 allowed for detailed characterization of their structural features, redox properties, stabilities, and interactions with cardiolipin (CL)-containing liposomes. Using a variety of spectroscopic tools, we show that CYC-2.1 and CYC-2.2 adopt a globular α-helical fold with His/Met heme ligation. The longer CYC-2.2 has a lower thermodynamic stability than CYC-2.1 and lacks His residues to misligate to the heme in the protein's denatured state. Both C. elegans proteins bind to CL-containing liposomes, and these interactions promote the proteins' peroxidase activity but to a much greater degree for CYC-2.2. Dye-to-heme distance distributions from time-resolved fluorescence resonance energy transfer in bimane-labeled CYC-2.1 and CYC-2.2 revealed similar populations of extended and compact conformers for CL-bound proteins, suggesting that their distinct peroxidase activities in the presence of CL arise from differences in the local heme environments for the two polypeptide ensembles. Without inhibition from His misligation, a less stable and more prone to unfolding CYC-2.2 allows for better access of substrates to the heme and thus exhibits higher peroxidase activity. Similar features of the conformational ensembles of CYC-2.1 and CYC-2.2 to those of mammalian cyt c suggest that C. elegans proteins, particularly the former, could serve as useful models for examining the mechanism of cyt c-CL interactions in live organisms.
Authors:
Amber J Vincelli; Danielle S Pottinger; Fangfang Zhong; Jonas Hanske; Stéphane G Rolland; Barbara Conradt; Ekaterina V Pletneva
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2013-01-16
Journal Detail:
Title:  Biochemistry     Volume:  52     ISSN:  1520-4995     ISO Abbreviation:  Biochemistry     Publication Date:  2013 Jan 
Date Detail:
Created Date:  2013-01-29     Completed Date:  2013-03-25     Revised Date:  2014-01-30    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  United States    
Other Details:
Languages:  eng     Pagination:  653-66     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Caenorhabditis elegans Proteins / biosynthesis,  chemistry*
Cardiolipins / chemistry*
Conserved Sequence
Cytochromes c / biosynthesis,  chemistry*
Escherichia coli
Fluorescence Resonance Energy Transfer
Guaiacol / chemistry
Heme / chemistry
Horses
Kinetics
Liposomes / chemistry
Models, Molecular
Molecular Sequence Data
Oxidation-Reduction
Peroxidases / biosynthesis,  chemistry*
Protein Binding
Protein Stability
Protein Structure, Secondary
Protein Structure, Tertiary
Protein Unfolding
Recombinant Proteins / biosynthesis,  chemistry
Sequence Analysis, Protein
Spectrophotometry, Ultraviolet
Thermodynamics
Grant Support
ID/Acronym/Agency:
GM069950/GM/NIGMS NIH HHS; GM076651/GM/NIGMS NIH HHS; GM098502/GM/NIGMS NIH HHS; R01 GM098502/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Caenorhabditis elegans Proteins; 0/Cardiolipins; 0/Liposomes; 0/Recombinant Proteins; 42VZT0U6YR/Heme; 6JKA7MAH9C/Guaiacol; 9007-43-6/Cytochromes c; EC 1.11.1.-/Peroxidases
Comments/Corrections

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