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
![[img]](http://spectrum.library.concordia.ca/style/images/fileicons/application_pdf.png)
| PDF (published version) 396Kb |
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 |
| References: | Boulanger, P., le Maire, M., Bonhivers, M., Dubois, S., Desmadril, M., and Letellier, L. (1996) Biochemistry 35, 14216–14224
Braun, V., Hantke, K., and Köster, W. (1998) in Metal Ions in Biological Systems (Sigel, A., and Sigel, H., eds) Vol. 35, pp. 67–145, Marcel Dekker, Inc., New York Braun, V., and Braun, M. (2002) Curr. Opin. Microbiol. 5, 194–201 Buchanan, S. K., Smith, B. S., Venkatramani, L., Xia, D., Esser, L., Palnitkar, M., Chakraborty, R., van der Helm, D., and Deisenhofer, J. (1999) Nat. Struct. Biol. 6, 56–63 Cadieux, N., and Kadner, R. J. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 10673–10678 Cadieux, N., Bradbeer, C., and Kadner, R. J. (2000) J. Bacteriol. 182, 5954–5961 Chang, C., Mooser, A., Pluckthun, A., and Wlodawer, A. (2001) J. Biol. Chem. 276, 27535–27540 Chimento, D. P., Kadner, R. J., and Wiener, M. C. (2003) J. Mol. Biol. 332, 999–1014 Chimento, D. P., Mohanty, A. K., Kadner, R. J., and Wiener, M. C. (2003) Nat. Struct. Biol. 10, 394–401 De Crescenzo, G., Grothe, S., Lortie, R., Debanne, M. T., and O'Connor-McCourt, M. (2000) Biochemistry 39, 9466–9476 De Crescenzo, G., Grothe, S., Zwaagstra, J., Tsang, M., and O'Connor-McCourt, M. D. (2001) J. Biol. Chem. 276, 29632–29643 Evans, J. S., Levine, B. A., Trayer, I. P., Dorman, C. J., and Higgins, C. F. (1986) FEBS Lett. 208, 211–216 Ferguson, A. D., Coulton, J. W., Diederichs, K., and Welte, W. (2001) in Handbook of Metalloproteinsin (Messerschmidt, A., Huber, R., Poulos T., and Wieghardt, K., eds) pp. 834–849, John Wiley & Sons, Ltd., Chichester Ferguson, A. D., Hofmann, E., Coulton, J. W., Diederichs, K., and Welte, W. (1998) Science 282, 2215–2220 Ferguson, A. D., Chakraborty, R., Smith, B. s., Esser, L., van der Helm, D., and Deisenhofer, J. (2002) Science 295, 1715–1719 Ferguson, A. D., Breed, J., Diederichs, K., Welte, W., and Coulton, J. W. (1998) Protein Sci. 7, 1636–1638 Gunter, K., and Braun, V. (1990) FEBS Lett. 274, 85–88 Johnsson, B., Lofas, S., and Lindquist, G. (1991) Anal. Biochem. 198, 268–277 Larsen, R. A., Letain, T. E., and Postle, K. (2003) Mol. Microbiol. 49, 211–218 Locher, K. P., Rees, B., Koebnik, R., Mitschler, A., Moulinier, L., Rosenbusch, J. P., and Moras, D. (1998) Cell 95, 771–778 Moeck, G. S., Tawa, P., Xiang, H., Ismail, A. A., Turnbull, J. L., and Coulton, J. W. (1996) Mol. Microbiol. 22, 459–471 Moeck, G. S., and Coulton, J. W. (1998) Mol. Microbiol. 28, 675–681 Moeck, G. S., Coulton, J. W., and Postle, K. (1997) J. Biol. Chem. 272, 28391–28397 Moeck, G. S., and Letellier, L. (2001) J. Bacteriol. 183, 2755–2764 Perkins, S. J. (1986) Eur. J. Biochem. 157, 169–180 Postle, K., and Kadner, R. J. (2003) Mol. Microbiol. 49, 869–882 Rich, R. L., and Myszka, D. G. (2000) Curr. Opin. Biotechnol. 11, 54–61 Sauter, A., Howard, S. P., and Braun, V. (2003) J. Bacteriol. 185, 5747–5754 Schoffler, H., and Braun, V. (1989) Mol. Gen. Genet. 217, 378–383 Schuck, P. (2000) Biophys. J. 78, 1606–1619 Skare, J. T., Ahmer, B. M., Seachord, C. L., Darveau, R. P., and Postle, K. (1993) J. Biol. Chem. 268, 16302–16308 Zhai, Y. F., Heijne, W., and Saier, M. H., Jr. (2003) Biochim. Biophys. Acta 1614, 201–210 |
Repository Staff Only: item control page
