Carter, David M. and Miousse, Isabelle R. and Gagnon, Jean-Nicolas and Martinez, Éric and Clements, Abigail and Lee, Jongchan and Hancock, Mark A. and Gagnon, Hubert and Pawelek, Peter D. and Coulton, James W. (2006) Interactions between TonB from Escherichia coli and the Periplasmic Protein FhuD. Journal of Biological Chemistry, 281 (46). pp. 35413-35424. ISSN 0021-9258
![[img]](http://spectrum.library.concordia.ca/style/images/fileicons/application_pdf.png)
| PDF (published version) 282Kb |
Official URL: http://www.jbc.org/cgi/content/abstract/281/46/354...
Abstract
For uptake of ferrichrome into bacterial cells, FhuA, a TonB-dependent outer membrane receptor of Escherichia coli, is required. The periplasmic protein FhuD binds and transfers ferrichrome to the cytoplasmic membrane-associated permease FhuB/C. We exploited phage display to map protein-protein interactions in the E. coli cell envelope that contribute to ferrichrome transport. By panning random phage libraries against TonB and against FhuD, we identified interaction surfaces on each of these two proteins. Their interactions were detected in vitro by dynamic light scattering and indicated a 1:1 TonB-FhuD complex. FhuD residue Thr-181, located within the siderophorebinding site and mapping to a predicted TonB-interaction surface, was mutated to cysteine. FhuD T181C was reacted with two thiol-specific fluorescent probes; addition of the siderophore ferricrocin quenched fluorescence emissions of these conjugates. Similarly, quenching of fluorescence from both probes confirmed binding of TonB and established an apparent KD of 300 nM. Prior saturation of the siderophorebinding site of FhuD with ferricrocin did not alter affinity of TonB for FhuD. Binding, further characterized with surface plasmon resonance, indicated a higher affinity complex with KD values in the low nanomolar range. Addition of FhuD to a preformed TonB-FhuA complex resulted in formation of a ternary complex. These observations led us to propose a novel mechanism in which TonB acts as a scaffold, directing FhuD to regions within the periplasm where it is poised to accept and deliver siderophore.
| Divisions: | Concordia University > Faculty of Arts and Science > Chemistry and Biochemistry |
|---|---|
| Item Type: | Article |
| Refereed: | Yes |
| Authors: | Carter, David M. and Miousse, Isabelle R. and Gagnon, Jean-Nicolas and Martinez, Éric and Clements, Abigail and Lee, Jongchan and Hancock, Mark A. and Gagnon, Hubert and Pawelek, Peter D. and Coulton, James W. |
| Journal or Publication: | Journal of Biological Chemistry |
| Date: | November 2006 |
| ID Code: | 6341 |
| Deposited By: | KUMIKO VEZINA |
| Deposited On: | 14 Sep 2009 15:05 |
| Last Modified: | 08 Dec 2010 18:56 |
| References: | Borths, E. L., Poolman, B., Hvorup, R. N., Locher, K. P., and Rees, D. C. (2005) Biochemistry 44, 16301-16309
Borths, E. L., Locher, K. P., Lee, A. T., and Rees, D. C. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 16642-16647 Carter, D. M., Gagnon, J. N., Damlaj, M., Mandava, S., Makowski, L., Rodi, D. J., Pawelek, P. D., and Coulton, J. W. (2006) J. Mol. Biol. 357, 236-251 Chang, C., Mooser, A., Plückthun, A., and Wlodawer, A. (2001) J. Biol. Chem. 276, 27535-27540 Clarke, T. E., Ku, S. Y., Dougan, D. R., Vogel, H. J., and Tari, L. W. (2000) Nat. Struct. Biol. 7, 287-291 Clarke, T. E., Braun, V., Winkelmann, G., Tari, L. W., and Vogel, H. J. (2002) J. Biol. Chem. 277, 13966-13972 Ferguson, A. D., Hofmann, E., Coulton, J. W., Diederichs, K., and Welte, W. (1998) Science 282, 2215-2220 Karpowich, N. K., Huang, H. H., Smith, P. C., and Hunt, J. F. (2003) J. Biol. Chem. 278, 8429-8434 Ködding, J., Killig, F., Polzer, P., Howard, S. P., Diederichs, K., and Welte, W. (2005) J. Biol. Chem. 280, 3022-3028 Köster, W., and Braun, V. (1990) J. Biol. Chem. 265, 21407-21410 Krewulak, K. D., Peacock, R. S., and Vogel, H. J. (2004) in Iron Transport in Bacteria (Crosa, J. H., Mey, A. R., and Payne, S. M., eds) pp. 113-129, American Society for Microbiology, Washington, D. C. Krewulak, K. D., Shepherd, C. M., and Vogel, H. J. (2005) Biometals 18, 375-386 Khursigara, C. M., De Crescenzo, G., Pawelek, P. D., and Coulton, J. W. (2004) J. Biol. Chem. 279, 7405-7412 Khursigara, C. M., De Crescenzo, G., Pawelek, P. D., and Coulton, J. W. (2005) Biochemistry 44, 3441-3453 Khursigara, C. M., De Crescenzo, G., Pawelek, P. D., and Coulton, J. W. (2005) Protein Sci. 14, 1266-1273 Lamb, A. L., Torres, A. S., O'Halloran, T. V., and Rosenzweig, A. C. (2000) Biochemistry 39, 14720-14727 Lawrence, M. C., Pilling, P. A., Epa, V. C., Berry, A. M., Ogunniyi, A. D., and Paton, J. C. (1998) Structure (Lond.) 6, 1553-1561 Lebowitz, J., Lewis, M. S., and Schuck, P. (2002) Protein Sci. 11, 2067-2079 Lee, Y. H., Deka, R. K., Norgard, M. V., Radolf, J. D., and Hasemann, C. A. (1999) Nat. Struct. Biol. 6, 628-633 Locher, K. P., Rees, B., Koebnik, R., Mitschler, A., Moulinier, L., Rosen-busch, J. P., and Moras, D. (1998) Cell 95, 771-778 Mademidis, A., Killmann, H., Kraas, W., Flechsler, I., Jung, G., and Braun, V. (1997) Mol. Microbiol. 26, 1109-1123 Mader, C., Huber, C., Moll, D., Sleytr, U. B., and Sára, M. (2004) J. Bacteriol. 186, 1758-1768 Mandava, S., Makowski, L., Devarapalli, S., Uzubell, J., and Rodi, D. J. (2004) Proteomics 4, 1439-1460 Moeck, G. S., and Letellier, L. (2001) J. Bacteriol. 183, 2755-2764 Myszka, D. G. (1999) J. Mol. Recognit. 12, 279-284 Nakajima, H., Cocquerel, L., Kiyokawa, N., Fujimoto, J., and Levy, S. (2005) Biochem. Biophys. Res. Commun. 328, 1091-1100 Pawelek, P. D., Croteau, N., Ng-Thow-Hing, C., Khursigara, C. M., Moi-seeva, N., Allaire, M., and Coulton, J. W. (2006) Science 312, 1399-1402 Peacock, R. S., Andrushchenko, V. V., Demcoe, A. R., Gehmlich, M., Lu, L. S., Herrero, A. G., and Vogel, H. J. (2006) Biometals 19, 127-142 Peacock, R. S., Weljie, A. M., Howard, S. P., Price, F. D., and Vogel, H. J. (2005) J. Mol. Biol. 345, 1185-1197 Rohrbach, M. R., Braun, V., and Köster, W. (1995) J. Bacteriol. 177, 7186-7193 Rohrbach, M. R., Paul, S., and Köster, W. (1995) Mol. Gen. Genet. 248, 33-42 Schuck, P. (2000) Biophys. J. 78, 1606-1619 Schultz-Hauser, G., Köster, W., Schwarz, H., and Braun, V. (1992) J. Bacteriol. 174, 2305-2311 Shultis, D. D., Purdy, M. D., Banchs, C. N., and Wiener, M. C. (2006) Science 312, 1396-1399 Sprencel, C., Cao, Z., Qi, Z., Scott, D. C., Montague, M. A., Ivanoff, N., Xu, J., Raymond, K. M., Newton, S. M., and Klebba, P. E. (2000) J. Bacteriol. 182, 5359-5364 Yang, Y. R., and Schachman, H. K. (1996) Biophys. Chem. 59, 289-297 Wandersman, C., and Delepelaire, P. (2004) Annu. Rev. Microbiol. 58, 611-647 |
Repository Staff Only: item control page
