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Enhanced Binding of TonB to a Ligand-loaded Outer Membrane Receptor: ROLE OF THE OLIGOMERIC STATE OF TonB IN FORMATION OF A FUNCTIONAL FhuA·TonB COMPLEX

Title:

Enhanced Binding of TonB to a Ligand-loaded Outer Membrane Receptor: ROLE OF THE OLIGOMERIC STATE OF TonB IN FORMATION OF A FUNCTIONAL FhuA·TonB COMPLEX

Khursigara, Cezar M. and De Crescenzo, Gregory and Pawelek, Peter D. and Coulton, James W. (2004) Enhanced Binding of TonB to a Ligand-loaded Outer Membrane Receptor: ROLE OF THE OLIGOMERIC STATE OF TonB IN FORMATION OF A FUNCTIONAL FhuA·TonB COMPLEX. Journal of Biological Chemistry, 279 (9). pp. 7405-7412. ISSN 0021-9258

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Official URL: http://www.jbc.org/cgi/content/abstract/279/9/7405

Abstract

The ferric hydroxymate uptake (FhuA) receptor from Escherichia coli facilitates transport of siderophores ferricrocin and ferrichrome and siderophore-antibiotic conjugates such as albomycin and rifamycin CGP 4832. FhuA is also the receptor for phages T5, T1, 80, UC-1, for colicin M and for the antimicrobial peptide microcin MccJ21. Energy for transport is provided by the cytoplasmic membrane complex TonB·ExbB·ExbD, which uses the proton motive force of the cytoplasmic membrane to transduce energy to the outer membrane. To accomplish energy transfer, TonB contacts outer membrane receptors. However, the stoichiometry of TonB· receptor complexes and their sites of interaction remain uncertain. In this study, analyses of FhuA interactions with two recombinant TonB proteins by analytical ultracentrifugation revealed that TonB forms a 2:1 complex with FhuA. The presence of the FhuA-specific ligand ferricrocin enhanced the amounts of complex but is not essential for its formation. Surface plasmon resonance experiments demonstrated that FhuA·TonB interactions are multiple and have apparent affinities in the nanomolar range. TonB also possesses two distinct binding regions: one in the C terminus of the protein, for which binding to FhuA is ferricrocin-independent, and a higher affinity region outside the C terminus, for which ferricrocin enhances interactions with FhuA. Together these experiments establish that FhuA·TonB interactions are more intricate than originally predicted, that the TonB·FhuA stoichiometry is 2:1, and that ferricrocin modulates binding of FhuA to TonB at regions outside the C-terminal domain of TonB.

Divisions:Concordia University > Faculty of Arts and Science > Chemistry and Biochemistry
Item Type:Article
Refereed:Yes
Authors:Khursigara, Cezar M. and De Crescenzo, Gregory and Pawelek, Peter D. and Coulton, James W.
Journal or Publication:Journal of Biological Chemistry
Date:February 2004
ID Code:6343
Deposited By:KUMIKO VEZINA
Deposited On:14 Sep 2009 15:43
Last Modified:08 Dec 2010 18:56
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