Copurification of the FpvA ferric pyoverdin receptor of Pseudomonas aeruginosa with its iron-free ligand: implications for siderophore-mediated iron transport
| Autor: | Schalk Ij, Prome D, Poole K, Abdallah Ma, Van Dorsselaer A, Pattus F, Pavel Kyslík |
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| Přispěvatelé: | Institut Gilbert-Laustriat : Biomolécules, Biotechnologie, Innovation Thérapeutique, Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Laboratory of Enzyme Technology, Czech Academy of Sciences [Prague] (CAS), Institut de pharmacologie et de biologie structurale (IPBS), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Laboratoire de Spectrométrie de Masse BioOrganique [Strasbourg] (LSMBO), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Department of Microbiology and Immunology (D MICROBIOLOGY IMMUNOLOGY), Queen's University [Kingston, Canada], Czech Academy of Sciences [Prague] (ASCR), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Spectrométrie de masse Bio-organique, UMR CNRS-ULP, Queen's University [Kingston], Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 1999 |
| Předmět: |
Circular dichroism
Siderophore Siderophores MESH: Amino Acid Sequence Ligands 7. Clean energy Biochemistry Copurification MESH: Circular Dichroism FepA MESH: Ligands MESH: Endopeptidases MESH: Bacterial Proteins 0303 health sciences Chemistry Circular Dichroism Hydrolysis Ligand (biochemistry) Transport protein MESH: Oligopeptides MESH: Pseudomonas aeruginosa Pseudomonas aeruginosa Bacterial outer membrane Oligopeptides MESH: Hydrolysis medicine.drug Bacterial Outer Membrane Proteins Protein Binding MESH: Pigments Biological MESH: Biological Transport Molecular Sequence Data Iron Chelating Agents 03 medical and health sciences Bacterial Proteins Endopeptidases medicine MESH: Protein Binding [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Amino Acid Sequence MESH: Siderophores 030304 developmental biology MESH: Molecular Sequence Data 030306 microbiology MESH: Bacterial Outer Membrane Proteins Biological Transport Pigments Biological biochemical phenomena metabolism and nutrition MESH: Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization MESH: Iron Chelating Agents Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Ferric |
| Zdroj: | Biochemistry Biochemistry, American Chemical Society, 1999, 38 (29), pp.9357-65. ⟨10.1021/bi990421x⟩ |
| ISSN: | 0006-2960 1520-4995 |
| DOI: | 10.1021/bi990421x⟩ |
| Popis: | The Pseudomonas aeruginosa FpvA receptor is a TonB-dependent outer membrane transport protein that catalyzes uptake of ferric pyoverdin across the outer membrane. Surprisingly, FpvA expressed in P. aeruginosa grown in an iron-deficient medium copurifies with a ligand X that we have characterized by UV, fluorescence, and mass spectrometry as being iron-free pyoverdin (apo-PaA). PaA was absent from FpvA purified from a PaA-deficient P. aeruginosa strain. The properties of ligand binding in vitro revealed very similar affinities of apo-PaA and ferric-PaA to FpvA. Fluorescence resonance energy transfer was used to study in vitro the formation of the FpvA-PaA-Fe complex in the presence of PaA-Fe or citrate-Fe. The circular dichroism spectrum of FpvA indicated a 57% beta-structure content typical of porins and in agreement with the 3D structures of the siderophore receptors FhuA and FepA. In the absence of the protease's inhibitors, a truncated form of FpvA lacking 87 amino acids at its N-terminus was purified. This truncated form still bound PaA, and its beta-sheet content was conserved. This N-terminal region displays significant homology to the N-terminal periplasmic extensions of FecA from Escherichia coli and PupB from Pseudomonas putida, which were previously shown to be involved in signal transduction. This suggests a similar function for FpvA. The mechanism of iron transport in P. aeruginosa via the pyoverdin pathway is discussed in the light of all these new findings. |
| Databáze: | OpenAIRE |
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