Abstract
FpvA is the primary outer membrane transporter required for iron acquisition via the siderophore pyoverdine (Pvd) in Pseudomonas aeruginosa. FpvA, like other ferrisiderophore transporters, consists of a membrane-spanning β-barrel occluded by a plug domain. The β-strands of the barrel are connected by large extracellular loops and periplasmic turns. Like some other TonB-dependent transporters, FpvA has a periplasmic domain involved in a signalling cascade that regulates expression of genes required for ferrisiderophore transport. Here, the structures of FpvA in different loading states are analysed in light of mutagenesis data. This analysis highlights the roles of different protein domains in Pvd-Fe uptake and the signalling cascade and reveals a strong correlation between Pvd-Fe transport and activation of the signalling cascade. It is likely that conclusions drawn for FpvA will be relevant to other TonB-dependent ferrisiderophore transport and signalling proteins.
Similar content being viewed by others
References
Adams H, Zeder-Lutz G, Greenwald J, Schalk IJ, Célia H, Pattus F (2006) Interaction of TonB with outer membrane receptor FpvA of Pseudomonas aeruginosa. J Bacteriol 188:5752–5761. doi:10.1128/JB.00435-06
Beare PA, For RJ, Martin LW, Lamont IL (2003) Siderophore-mediated cell signalling in Pseudomonas aeruginosa: divergent pathways regulate virulence factor production and siderophore receptor synthesis. Mol Microbiol 47:195–207. doi:10.1046/j.1365-2958.2003.03288.x
Bos C, Lorenzen D, Braun V (1998) Specific in vivo labeling of cell surface-exposed protein loops: reactive cysteines in the predicted gating loop mark a ferrichrome binding site and a ligand-induced conformational change of the Escherichia coli FhuA protein. J Bacteriol 180:605–613
Boukhalfa H, Crumbliss AL (2002) Chemical aspects of siderophore mediated iron transport. Biometals 15:325–339. doi:10.1023/A:1020218608266
Braun V, Braun M (2002) Iron transport and signaling in Escherichia coli. FEBS Lett 529:78–85. doi:10.1016/S0014-5793(02)03185-X
Brillet K, Journet L, Celia H, Paulus L, Stahl A, Pattus F, Cobessi D (2007) A β-strand lock-exchange for signal transduction in TonB-dependent transducers on the basis of a common structural motif. Structure 15:1383–1391. doi:10.1016/j.str.2007.08.013
Cao Z, Warfel P, Newton SM, Klebba PE (2003) Spectroscopic observations of ferric enterobactin transport. J Biol Chem 278:1022–1028. doi:10.1074/jbc.M210360200
Cobessi D, Célia H, Folschweiller N, Schalk IJ, Abdallah MA, Pattus F (2005) The crystal structure of the pyoverdin outer membrane receptor FpvA from Pseudomonas aeruginosa at 3.6 Å resolution. J Mol Biol 34:121–134. doi:10.1016/j.jmb.2005.01.021
Eisenhauer HA, Shames S, Pawelek PD, Coulton JW (2005) Siderophore transport through Escherichia coli outer membrane receptor FhuA with disulfide-tethered cork and barrel domains. J Biol Chem 280:30574–30580. doi:10.1074/jbc.M506708200
Endriss F, Braun M, Killmann H, Braun V (2003) Mutant analysis of the Escherichia coli FhuA protein reveals sites of FhuA activity. J Bacteriol 185:4683–4692. doi:10.1128/JB.185.16.4683-4692.2003
Enz S, Mahren S, Stroeher UH, Braun V, Ochs M, Veitinger S, Kim I, Welz D, Angerer A, Hantke K (2000) Surface signaling in ferric citrate transport gene induction: interaction of the FecA, FecR, and FecI regulatory proteins. J Bacteriol 182:637–646. doi:10.1128/JB.182.3.637-646.2000
Ferguson AD, Deisenhofer J (2004) Metal import through microbial membranes. Cell 116:15–24. doi:10.1016/S0092-8674(03)01030-4
Ferguson AD, Hofmann E, Coulton JW, Diederichs K, Welte W (1998) Siderophore-mediated iron transport: crystal structure of FhuA with bound lipopolysaccharide. Science 282:2215–2220. doi:10.1126/science.282.5397.2215
Ferguson AD, Chakraborty R, Smith BS, Esser L, van der Helm D, Deisenhofer J (2002) Structural basis of gating by the outer membrane transporter FecA. Science 295:1715–1719. doi:10.1126/science.1067313
Ferguson AD, Amezcua CA, Halabi NM, Chelliah Y, Rosen MK, Ranganathan R, Deisenhofer J (2007) Signal transduction pathway of TonB-dependent transporters. Proc Natl Acad Sci USA 104:513–518. doi:10.1073/pnas.0609887104
Folschweiller N, Gallay J, Vincent M, Abdallah MA, Pattus F, Schalk IJ (2002) The interaction between pyoverdin and its outer membrane receptor in Pseudomonas aeruginosa leads to different conformers: a time-resolved fluorescence study. Biochemistry 41:14591–14601. doi:10.1021/bi0259711
Garcia-Herrero A, Vogel HJ (2005) Nuclear magnetic resonance solution structure of the periplasmic signalling domain of the TonB-dependent outer membrane transporter FecA from Escherichia coli. Mol Microbiol 58:1226–1237
Gumbart J, Wiener MC, Tajkhorshid E (2007) Mechanics of force propagation in TonB-dependent outer membrane transport. Biophys J 93:496–504. doi:10.1529/biophysj.107.104158
Hantke K (2005) Bacterial zinc uptake and regulators. Curr Opin Microbiol 8:196–202. doi:10.1016/j.mib.2005.02.001
James HE, Beare PA, Martin LW, Lamont IL (2005) Mutational analysis of a bifunctional ferrisiderophore receptor and signal-transducing protein from Pseudomonas aeruginosa. J Bacteriol 187:4514–4520. doi:10.1128/JB.187.13.4514-4520.2005
Lamont IL, Martin LW (2003) Identification and characterization of novel pyoverdine synthesis genes in Pseudomonas aeruginosa. Microbiology 149:833–842. doi:10.1099/mic.0.26085-0
Lamont IL, Beare PA, Ochsner U, Vasil AI, Vasil ML (2002) Siderophore-mediated signaling regulates virulence factor production in Pseudomonas aeruginosa. Proc Natl Acad Sci USA 99:7072–7077. doi:10.1073/pnas.092016999
Locher KP, Rees B, Koebnik R, Mitschler A, Moulinier L, Rosenbusch JP, Moras D (1998) Transmembrane signaling across the ligand-gated FhuA receptor: crystal structures of free and ferrichrome-bound states reveal allosteric changes. Cell 95:771–778. doi:10.1016/S0092-8674(00)81700-6
Ma L, Kaserer W, Annamalai R, Scott DC, Jin B, Jiang X, Xiao Q, Maymani H, Massis LM, Ferreira LC, Newton SM, Klebba PE (2007) Evidence of ball-and-chain transport of ferric enterobactin through FepA. J Biol Chem 282:397–406. doi:10.1074/jbc.M605333200
Nader M, Dobbelaere W, Vincent M, Journet L, Adams C, Cobessi D, Gallay J, Schalk IJ (2007) Identification of residues of FpvA involved in the different steps of Pvd-Fe uptake in Pseudomonas aeruginosa. Biochemistry 46:11707–11717. doi:10.1021/bi700997w
Neilands JB (1995) Siderophores: structure and function of microbial iron transport compounds. J Biol Chem 270:26723–26726
Pawelek PD, Croteau N, Ng-Thow-Hing C, Khursigara CM, Moiseeva N, Allaire M, Coulton JW (2006) Structure of TonB in complex with FhuA, E. coli outer membrane receptor. Science 312:1399–1402. doi:10.1126/science.1128057
Poole K, Neshat S, Krebes K, Heinrichs DE (1993) Cloning and nucleotide sequence analysis of the ferripyoverdine receptor gene fpvA of Pseudomonas aeruginosa. J Bacteriol 175:4597–4604
Postle K, Larsen RA (2007) TonB-dependent energy transduction between outer and cytoplasmic membranes. Biometals 20:453–465. doi:10.1007/s10534-006-9071-6
Redly GA, Poole K (2005) FpvIR control of fpvA ferric pyoverdine receptor gene expression in Pseudomonas aeruginosa: demonstration of an interaction between FpvI and FpvR and identification of mutations in each compromising this interaction. J Bacteriol 187:5648–5657. doi:10.1128/JB.187.16.5648-5657.2005
Sauter A, Braun V (2004) Defined inactive FecA derivatives mutated in functional domains of the outer membrane transport and signaling protein of Escherichia coli K-12. J Bacteriol 186:5303–5310. doi:10.1128/JB.186.16.5303-5310.2004
Schons V, Atkinson RA, Dugave C, Graff R, Mislin GL, Rochet L, Hennard C, Kieffer B, Abdallah MA, Schalk IJ (2005) The structure–activity relationship of ferric pyoverdine bound to its outer membrane transporter: implications for the mechanism of iron uptake. Biochemistry 44:14069–14079. doi:10.1021/bi051155s
Scott DC, Newton SM, Klebba PE (2002) Surface loop motion in FepA. J Bacteriol 184:4906–4911. doi:10.1128/JB.184.17.4906-4911.2002
Shen J, Meldrum A, Poole K (2002) FpvA receptor involvement in pyoverdine biosynthesis in Pseudomonas aeruginosa. J Bacteriol 184:3268–3275. doi:10.1128/JB.184.12.3268-3275.2002
Shen JS, Geoffroy V, Neshat S, Jia Z, Meldrum A, Meyer JM, Poole K (2005) FpvA-mediated ferric pyoverdine uptake in Pseudomonas aeruginosa: identification of aromatic residues in FpvA implicated in ferric pyoverdine binding and transport. J Bacteriol 187:8511–8515. doi:10.1128/JB.187.24.8511-8515.2005
Shultis DD, Purdy MD, Banchs CN, Wiener MC (2006) Outer membrane active transport: structure of the BtuB:TonB complex. Science 312:1396–1399. doi:10.1126/science.1127694
Visca P (2004) Iron regulation and siderophore signalling in virulence by Pseudomonas aeruginosa. In: Juan-Luis R (ed) Pseudomonas, vol 2. Kluwer Academic/Plenum Publishers, New York, pp 69–123
Visca P, Leoni L, Wilson MJ, Lamont IL (2002) Iron transport and regulation, cell signalling and genomics: lessons from Escherichia coli and Pseudomonas. Mol Microbiol 45:1177–1190. doi:10.1046/j.1365-2958.2002.03088.x
Wirth C, Meyer-Klaucke W, Pattus F, Cobessi D (2007) From the periplasmic signaling domain to the extracellular face of an outer membrane signal transducer of Pseudomonas aeruginosa: crystal structure of the ferric pyoverdine outer membrane receptor. J Mol Biol 68:398–406. doi:10.1016/j.jmb.2007.02.023
Yue WW, Grizot S, Buchanan SK (2003) Structural evidence for iron-free citrate and ferric citrate binding to the TonB-dependent outer membrane transporter FecA. J Mol Biol 332:353–368. doi:10.1016/S0022-2836(03)00855-6
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Schalk, I.J., Lamont, I.L. & Cobessi, D. Structure–function relationships in the bifunctional ferrisiderophore FpvA receptor from Pseudomonas aeruginosa . Biometals 22, 671–678 (2009). https://doi.org/10.1007/s10534-008-9203-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10534-008-9203-2