Document Detail


Interaction of TonB with the outer membrane receptor FpvA of Pseudomonas aeruginosa.
MedLine Citation:
PMID:  16885443     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Pyoverdine-mediated iron uptake by the FpvA receptor in the outer membrane of Pseudomonas aeruginosa is dependent on the inner membrane protein TonB1. This energy transducer couples the proton-electrochemical potential of the inner membrane to the transport event. To shed more light upon this process, a recombinant TonB1 protein lacking the N-terminal inner membrane anchor (TonB(pp)) was constructed. This protein was, after expression in Escherichia coli, purified from the soluble fraction of lysed cells by means of an N-terminal hexahistidine or glutathione S-transferase (GST) tag. Purified GST-TonB(pp) was able to capture detergent-solubilized FpvA, regardless of the presence of pyoverdine or pyoverdine-Fe. Targeting of the TonB1 fragment to the periplasm of P. aeruginosa inhibited the transport of ferric pyoverdine by FpvA in vivo, indicating an interference with endogenous TonB1, presumably caused by competition for binding sites at the transporter or by formation of nonfunctional TonB heterodimers. Surface plasmon resonance experiments demonstrated that the FpvA-TonB(pp) interactions have apparent affinities in the micromolar range. The binding of pyoverdine or ferric pyoverdine to FpvA did not modulate this affinity. Apparently, the presence of either iron or pyoverdine is not essential for the formation of the FpvA-TonB complex in vitro.
Authors:
Hendrik Adams; Gabrielle Zeder-Lutz; Isabelle Schalk; Franc Pattus; Hervé Celia
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of bacteriology     Volume:  188     ISSN:  0021-9193     ISO Abbreviation:  J. Bacteriol.     Publication Date:  2006 Aug 
Date Detail:
Created Date:  2006-08-03     Completed Date:  2006-09-18     Revised Date:  2013-06-07    
Medline Journal Info:
Nlm Unique ID:  2985120R     Medline TA:  J Bacteriol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5752-61     Citation Subset:  IM    
Affiliation:
ESBS UMR7175, Récepteurs et Protéines Membranaires, Rue Sebastien Brant, BP 10413 F-67412 Illkirch Cedex, France.
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MeSH Terms
Descriptor/Qualifier:
Bacterial Outer Membrane Proteins / genetics,  metabolism*
Bacterial Proteins / genetics,  metabolism*
Iron / metabolism
Membrane Proteins / genetics,  metabolism*
Oligopeptides / metabolism
Protein Binding
Pseudomonas aeruginosa / genetics,  metabolism*
Chemical
Reg. No./Substance:
0/Bacterial Outer Membrane Proteins; 0/Bacterial Proteins; 0/FpvA protein, Pseudomonas aeruginosa; 0/Membrane Proteins; 0/Oligopeptides; 0/tonB protein, Bacteria; 7439-89-6/Iron; 8062-00-8/pyoverdin
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