Document Detail

Spatial organization of the cytochrome b6-f complex within chloroplast thylakoid membranes.
MedLine Citation:
PMID:  3955028     Owner:  NLM     Status:  MEDLINE    
The spatial distribution of the chloroplast thylakoid protein complex comprised of cytochromes f and b-563, and the Rieske iron-sulfur protein (Cyt b6-f) has been controversial because of conflicting results obtained by different techniques. We have combined the following biochemical and immunochemical techniques to approach this question: (1) French press disruption of thylakoids, followed by repeated two-phase aqueous polymer partitioning to separate inside-out grana from right-side-out stroma membrane fragments; (2) electrophoretic analysis followed by the 3,3',5,5'-tetramethylbenzidine stain for cytochrome hemes; (3) electroblot analysis with anti-Cyt b6-f antibodies; (4) agglutination of membrane fragments with anti-Cyt b6-f antibodies; and (5) post-embedment thin-section immunolabeling of chemically fixed or ultrarapidly frozen chloroplasts with anti-Cyt b6-f antibodies. Our results indicate that the complex is present in both of the isolated membrane fragment populations in similar amounts, with the bulk of the immunoreactive sites exposed to the thylakoidal lumen. Direct immunolabeling of thin-sectioned chloroplasts resulted in localization of the complex throughout the thylakoids, without specialized compartmentation. These results provide both the temporal and spatial resolution necessary for accurate localization of the complex. We concur with models proposing distribution of Cyt b6-f throughout all thylakoid membranes.
D R Allred; L A Staehelin
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  849     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  1986 Apr 
Date Detail:
Created Date:  1986-05-07     Completed Date:  1986-05-07     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  94-103     Citation Subset:  IM    
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MeSH Terms
Chloroplasts / metabolism*,  ultrastructure
Cytochrome b Group / metabolism
Cytochrome b6f Complex
Cytochromes / metabolism*
Cytochromes f
Freeze Fracturing
Intracellular Membranes / metabolism,  ultrastructure
Microscopy, Electron
Plants / metabolism
Species Specificity
Grant Support
Reg. No./Substance:
0/Cytochrome b Group; 0/Cytochromes; 9035-40-9/Cytochrome b6f Complex; 9035-46-5/Cytochromes f

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