Mapping the interaction between the hemophore HasA and its outer membrane receptor HasR using CRINEPT-TROSY NMR spectroscopy
| Autor: | Anne Lecroisey, Célia Caillet-Saguy, Paola Turano, Ivano Bertini, Muriel Delepierre, Nadia Izadi-Pruneyre, Mario Piccioli |
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| Přispěvatelé: | Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), CERM and Deparment of Chemistry, Università degli Studi di Firenze = University of Florence [Firenze] (UNIFI), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Università degli Studi di Firenze = University of Florence (UniFI) |
| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
Models
Molecular Stereochemistry Protein Conformation Phosphorylcholine Molecular Sequence Data Receptors Cell Surface Heme 010402 general chemistry 01 natural sciences Biochemistry Micelle Catalysis Adduct 03 medical and health sciences chemistry.chemical_compound Colloid and Surface Chemistry Bacterial Proteins Amino Acid Sequence Receptor Nuclear Magnetic Resonance Biomolecular Micelles 030304 developmental biology 0303 health sciences [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] Membrane Proteins General Chemistry Nuclear magnetic resonance spectroscopy 0104 chemical sciences NMR spectra database chemistry Bacterial outer membrane Heme acquisition Apoproteins Carrier Proteins |
| Zdroj: | Journal of the American Chemical Society Journal of the American Chemical Society, American Chemical Society, 2009, 131 (5), pp.1736-44. ⟨10.1021/ja804783x⟩ Journal of the American Chemical Society, 2009, 131 (5), pp.1736-44. ⟨10.1021/ja804783x⟩ |
| ISSN: | 0002-7863 1520-5126 |
| DOI: | 10.1021/ja804783x⟩ |
| Popis: | The first step of heme acquisition by Gram-negative pathogenic bacteria through the so-called heme acquisition system, Has, requires delivery of the heme from the extracellular hemophore protein HasA to a specific outer membrane receptor, HasR. CRINEPT-TROSY NMR experiments in DPC micelles were here used to obtain information on the intermediate HasA-HasR complex in solution. A stable protein-protein adduct is detected both in the presence and in the absence of heme. Structural information on the complexed form of HasA is obtained from chemical shift mapping and statistical analysis of the spectral fingerprint of the protein NMR spectra obtained under different conditions. This approach shows the following: (i) only three different conformations are possible for HasA in solution: one for the isolated apoprotein, one for the isolated holoprotein, and one for the complexed protein, that is independent of the presence of the heme; (ii) the structure of the hemophore in the complex resembles the open conformation of the apoprotein; (iii) the surface contact area between HasA and HasR is independent of the presence of the heme, involving loop L1, loop L2, and the beta2-beta6 strands; (iv) upon complex formation the heme group is transferred from holoHasA to HasR. |
| Databáze: | OpenAIRE |
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