A group of Cu,Zn-superoxide dismutases from pathogenic bacteria is characterized by histidine-rich N-terminal extensions that are in a highly exposed and mobile conformation. This feature allows these proteins to be readily purified in a single step by immobilized metal affinity chromatography. The Cu,Zn-superoxide dismutases from both Haemophilus ducreyi and Haemophilus parainfluenzae display anomalous absorption spectra in the visible region due to copper binding at the N-terminal region. Reconstitution experiments of copper-free enzymes demonstrate that, under conditions of limited copper availability, this metal ion is initially bound at the N-terminal region and subsequently transferred to an active site. Evidence is provided for intermolecular pathways of copper transfer from the N-terminal domain of an enzyme subunit to an active site located on a distinct dimeric molecule. Incubation with EDTA rapidly removes copper bound at the N terminus but is much less effective on the copper ion bound at the active site. This indicates that metal binding by the N-terminal histidines is kinetically favored, but the catalytic site binds copper with higher affinity. We suggest that the histidine-rich N-terminal region constitutes a metal binding domain involved in metal uptake under conditions of metal starvation in vivo. Particular biological importance for this domain is inferred by the observation that its presence enhances the protection offered by periplasmic Cu,Zn-superoxide dismutase toward phagocytic killing.

Battistoni, A., Pacello, F., Mazzetti, A., Capo, C., Kroll, J., Langford, P., et al. (2001). A histidine-rich metal binding domain at the N terminus of Cu,Zn-superoxide dismutases from pathogenic bacteria. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 276(32), 30315-30325 [10.1074/jbc.M010527200].

A histidine-rich metal binding domain at the N terminus of Cu,Zn-superoxide dismutases from pathogenic bacteria

Battistoni A;Pacello F;Mazzetti AP;Capo C;Rotilio G
2001-01-01

Abstract

A group of Cu,Zn-superoxide dismutases from pathogenic bacteria is characterized by histidine-rich N-terminal extensions that are in a highly exposed and mobile conformation. This feature allows these proteins to be readily purified in a single step by immobilized metal affinity chromatography. The Cu,Zn-superoxide dismutases from both Haemophilus ducreyi and Haemophilus parainfluenzae display anomalous absorption spectra in the visible region due to copper binding at the N-terminal region. Reconstitution experiments of copper-free enzymes demonstrate that, under conditions of limited copper availability, this metal ion is initially bound at the N-terminal region and subsequently transferred to an active site. Evidence is provided for intermolecular pathways of copper transfer from the N-terminal domain of an enzyme subunit to an active site located on a distinct dimeric molecule. Incubation with EDTA rapidly removes copper bound at the N terminus but is much less effective on the copper ion bound at the active site. This indicates that metal binding by the N-terminal histidines is kinetically favored, but the catalytic site binds copper with higher affinity. We suggest that the histidine-rich N-terminal region constitutes a metal binding domain involved in metal uptake under conditions of metal starvation in vivo. Particular biological importance for this domain is inferred by the observation that its presence enhances the protection offered by periplasmic Cu,Zn-superoxide dismutase toward phagocytic killing.
2001
Pubblicato
Rilevanza internazionale
Articolo
Sì, ma tipo non specificato
Settore BIO/10 - BIOCHIMICA
English
Con Impact Factor ISI
Affinity chromatography; Bacteriology; Catalysis; Conformations; Dimers; Enzyme kinetics; Intermolecular pathways; Biochemistry; chaperone; copper; copper zinc superoxide dismutase; dimer; edetic acid; histidine; metal; protein; proteinase; superoxide dismutase; amino terminal sequence; animal cell; article; controlled study; enzyme active site; enzyme conformation; enzyme subunit; Haemophilus ducreyi; Haemophilus parainfluenzae; human; human cell; immobilized metal affinity chromatography; metal binding; mouse; nonhuman; phagocyte; priority journal; protein domain; protein purification; amino acid sequence; animal; binding site; chemistry; chromatography; dimerization; enzymology; Haemophilus influenzae; kinetics; macrophage; metabolism; molecular genetics; pathogenicity; phagocytosis; plasmid; protein binding; protein tertiary structure; sequence homology; time; Animalia; Haemophilus; Haemophilus ducreyi; Haemophilus parainfluenzae; Amino Acid Sequence; Animals; Binding Sites; Chromatography; Copper; Dimerization; Edetic Acid; Endopeptidases; Haemophilus ducreyi; Haemophilus influenzae; Histidine; Humans; Kinetics; Macrophages; Mice; Molecular Sequence Data; Phagocytosis; Plasmids; Protein Binding; Protein Structure, Tertiary; Proteins; Sequence Homology, Amino Acid; Superoxide Dismutase; Time Factors
Battistoni, A., Pacello, F., Mazzetti, A., Capo, C., Kroll, J., Langford, P., et al. (2001). A histidine-rich metal binding domain at the N terminus of Cu,Zn-superoxide dismutases from pathogenic bacteria. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 276(32), 30315-30325 [10.1074/jbc.M010527200].
Battistoni, A; Pacello, F; Mazzetti, A; Capo, C; Kroll, J; Langford, P; Sansone, A; Donnarumma, G; Valenti, P; Rotilio, G
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/55126
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