Crystal Structure of Reduced Bovine Erythrocyte Superoxide Dismutase at 1.9 Å Resolution

Rypniewski, W.; Mangani, Stefano; Bruni, B; Orioli, Pierluigi; Casati, Marco; Wilson, K.S.

doi:10.1006/jmbi.1995.0434

Cited by 89 publications

(76 citation statements)

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“…A main result concerns the rupture of the interaction between reduced Cu(I) and the His-63 bridging ligand, as deduced by NMR (10,(23)(24)(25), extended X-ray absorption fluorescence spectroscopy (26,27), and Raman spectroscopy (28 -30) for Cu,Zn-SOD in solution. Three-dimensional crystallographic structures of reduced Cu,Zn-SOD presented heterogeneity of the active center (16,(31)(32)(33), but a recent high resolution structure provided convincing evidence for three coordinated Cu(I) (34), in agreement with other spectroscopic data.…”

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confidence: 80%

“…Structural factors at the origin of this specific and efficient dismutase mechanism are not fully understood. Thorough analyses at the molecular level of structural changes that accompany copper reduction have been undertaken to detail the active site properties at the origin of the efficient superoxide dismutation and of the protonation events associated with its reduction into hydrogen peroxide (3,5,10,(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34). A main result concerns the rupture of the interaction between reduced Cu(I) and the His-63 bridging ligand, as deduced by NMR (10,(23)(24)(25), extended X-ray absorption fluorescence spectroscopy (26,27), and Raman spectroscopy (28 -30) for Cu,Zn-SOD in solution.…”

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confidence: 99%

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Long Distance Charge Redistribution Upon Cu,Zn-Superoxide Dismutase Reduction

Dupeyrat

Vidaud

Lorphelin

et al. 2004

Journal of Biological Chemistry

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Cu,Zn-superoxide dismutase (Cu,Zn-SOD) is a ubiquitous enzyme with an essential role in antioxidant defense. To better understand structural factors at the origin of the highly efficient superoxide dismutation mechanism, we analyzed the consequence of copper reduction on the electronic properties of the backbone and individual amino acids by using electrochemistry coupled to Fourier transform infrared spectroscopy. Comparison of data recorded with bovine erythrocyte and recombinant chloroplastic Cu,Zn-SOD from Lycopersicon esculentum, expressed as a functional tetramer in Escherichia coli and 14 N-or fully 15 N-labeled, demonstrated that the infrared changes were dominated by reorganizations of peptide bonds and histidine copper ligands. Two main infrared modes of histidine side chain, markers of metal coordination, were identified by using Cu-and Zn-methylimidazole models: the (C 4 C 5 ) at 1605-1594 cm ؊1 or Ϸ1586 cm ؊1 for N or N coordination, and the (C 5 N ) at Ϸ1113-1088 cm ؊1 . These modes, also identified in Cu,Zn-SOD by using 15 N labeling, showed that the electronic properties of the histidine N ligands of copper are mostly affected upon copper reduction. A striking conclusion of this work is that peptide groups from loops and ␤-sheet largely participate in charge redistribution upon copper reduction, and in contrast, electronic properties of polar and charged amino acids of the superoxide access channel remain unaffected. This is notably shown for the strictly conserved Arg-143 by site-directed mutagenesis on chloroplastic Cu,Zn-SOD. Charge compensation by the peptide backbone and preserved electronic properties of the superoxide access channel and docking site upon copper reduction may be the determinant factors for the high reaction kinetics of superoxide with both reduced and oxidized Cu,Zn-SOD.

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supporting

confidence: 80%

mentioning

confidence: 99%

Long Distance Charge Redistribution Upon Cu,Zn-Superoxide Dismutase Reduction

Dupeyrat

Vidaud

Lorphelin

et al. 2004

Journal of Biological Chemistry

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“…It has been suggested that this cysteine residue is sensitive to oxidative modification, which can result in enzyme inhibition. 24 Because the sheep CuZnSOD protein has not yet been analyzed, it is unclear whether the Cys55 equivalent residue in the ovine CuZnSOD enzyme would be sensitive to this type of oxidative inhibition.…”

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confidence: 99%

Increased Superoxide Generation Is Associated With Pulmonary Hypertension in Fetal Lambs

Brennan

Steinhorn

Wedgwood

et al. 2003

Circulation Research

215

189

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Abstract-Ligation of the ductus arteriosus in utero produces pulmonary hypertension and vascular remodeling in fetal and newborn lambs. However, the mechanisms producing these vascular changes are not well defined. Because reactive oxygen species (ROS) have been implicated as mediators of smooth muscle cell proliferation, we hypothesized that increased formation of ROS may be involved in the pathophysiology of pulmonary hypertension after in utero ductal ligation. Using ethidium fluorescence, we demonstrated an increase in superoxide levels after 9 days of ductal ligation compared with control lungs (PϽ0.05) that was localized to the adventitia and smooth muscle cells of hypertensive vessels. SOD-1 and SOD-2 protein levels and activities in lung, vein, and artery of hypertensive lambs were unchanged relative to controls after 2 days of ductal ligation. However, after 9 days, superoxide dismutase (SOD) activity was significantly decreased in arteries from ligated lambs without associated changes in SOD protein expression (PϽ0.05).Examination of NADPH oxidase expression as a potential source of the superoxide production indicated that the levels of p67 phox , a subunit of the NADPH oxidase complex, were significantly increased in the pulmonary arteries, but not veins, from the ligated lung as early as 2 days (PϽ0.05). Functional analyses demonstrated that reducing superoxide levels significantly increased the NO-mediated relaxation of pulmonary arteries isolated after 9 days, but not 2 days, of ductal ligation (PϽ0.05). These results suggest that increased NADPH oxidase expression may increase levels of superoxide in persistent pulmonary hypertension of the newborn lung tissue, and that increased superoxide blunts vascular relaxations to exogenous NO while stimulating smooth muscle cell growth. Key Words: reactive oxygen species Ⅲ vascular remodeling Ⅲ smooth muscle W ith initiation of ventilation and oxygenation at birth, pulmonary vascular resistance decreases and pulmonary blood flow increases. 1 In some neonates, this normal transition does not occur, resulting in persistent pulmonary hypertension of the newborn (PPHN). PPHN can occur idiopathically or as a complication of a variety of cardiorespiratory disorders, including hyaline membrane disease, asphyxia, meconium aspiration, and congenital diaphragmatic hernia. 2,3 In newborns that die with PPHN, there is an increase in both the thickness of the smooth muscle layer within small pulmonary arteries and an extension of this muscle to nonmuscular arteries. 4 There is also proliferation of adventitial tissue. 5 These structural changes indicate that in utero events have altered the pulmonary circulation. However, the mechanism underlying these changes is unclear. Surgical ligation or constriction of the ductus arteriosus in the fetal lamb several days before delivery at term results in PPHN after delivery. 6 -9 This lamb model closely resembles the human condition, with suprasystemic pulmonary arterial pressure and anatomic changes including muscularization o...

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“…The X-ray structure of the oxidized form has been available since 1982 for the bovine enzyme [6,7] and several other structures have become available [8][9][10][11][12][13][14][15][16][17][18]. Reduced state structures are also available although the picture is less clear-cut around the copper-binding site [19][20][21]. Certainties on the protonation of His63, which bridges Cu and Zn in the oxidized form but is protonated in the reduced form, come from 1 H NMR studies [22][23][24][25].…”

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confidence: 99%

The solution structure of reduced dimeric copper zinc superoxide dismutase. The structural effects of dimerization

Banci¹,

Bertini²,

Cramaro³

et al. 2002

Eur J Biochem

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The solution structure of homodimeric Cu 2 Zn 2 superoxide dismutase (SOD) of 306 aminoacids was determined on a 13 C, 15 N and 70% 2 H labeled sample. Two-thousand eighthundred and five meaningful NOEs were used, of which 96 intersubunit, and 115 dihedral angles provided a family of 30 conformers with an rmsd from the average of 0.78 ± 0.11 and 1.15 ± 0.09 Å for the backbone and heavy atoms, respectively. When the rmsd is calculated for each subunit, the values drop to 0.65 ± 0.09 and 1.08 ± 0.11 Å for the backbone and heavy atoms, respectively.The two subunits are identical on the NMR time scale, at variance with the X-ray structures that show structural differences between the two subunits as well as between different molecules in the unit cell. The elements of secondary structure, i.e. eight b sheets, are the same as in the X-ray structures and are well defined. The odd loops (I, III and V) are well resolved as well as loop II located at the subunit interface. On the contrary, loops IV and VI show some disorder. The residues of the active cavity are well defined whereas within the various subunits of the X-ray structure some are disordered or display different orientation in different X-ray structure determinations. The copper(I) ion and its ligands are well defined. This structure thus represents a well defined model in solution relevant for structure-function analysis of the protein. The comparison between the solution structure of monomeric mutants and the present structure shows that the subunit-subunit interactions increase the order in loop II. This has the consequences of inducing the structural and dynamic properties that are optimal for the enzymatic function of the wild-type enzyme. The regions 37-43 and 89-95, constituting loops III and V and the initial part of the b barrel and showing several mutations in familial amyotrophis lateral sclerosis (FALS)-related proteins have a quite extensive network of H-bonds that may account for their low mobility. Finally, the conformation of the key Arg143 residue is compared to that in the other dimeric and monomeric structures as well as in the recently reported structure of the CCS-superoxide dismutase (SOD) complex.

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Crystal Structure of Reduced Bovine Erythrocyte Superoxide Dismutase at 1.9 Å Resolution

Cited by 89 publications

References 0 publications

Long Distance Charge Redistribution Upon Cu,Zn-Superoxide Dismutase Reduction

Long Distance Charge Redistribution Upon Cu,Zn-Superoxide Dismutase Reduction

Increased Superoxide Generation Is Associated With Pulmonary Hypertension in Fetal Lambs

The solution structure of reduced dimeric copper zinc superoxide dismutase. The structural effects of dimerization

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