Crystallization and X-ray examination of bovine adrenodoxin

Marg, Andreas; Kuban, Ralf‐Jürgen; Behlke, Joachim; Dettmer, R; Ruckpaul, K

doi:10.1016/0022-2836(92)90235-c

Cited by 17 publications

(8 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although preliminary crystallographic data exists for bovine adrenodoxin (Marg et al, 1992), no X-ray structural information is available for this ferredoxin nor for any other vertebrate [2Fe-2S] ferredoxin. Structural information is available for some plant-type ferredoxins, as the crystal structures of the [2Fe-2S] ferredoxins from Spirulina platensis (Fukuyama et al, 1980;., 1990), Aphanothece sacrum (Tsukihara et al, 1990), Anabaena 7120 (Rypniewsky et al, 1991) and Euqiseturn arvense (Ikemizu et al, 1994) have been solved to 0.25, 0.22-, 0.25-and 0.18-nm resolution.…”

Section: Discussionmentioning

confidence: 99%

“…The final purification step was performed on a Sephadex G-50 column (Pharmacia) using 10 mM potassium phosphate, pH 7.4. Native bovine adrenodoxin for control fluorescence measurements was isolated from bovine adrenal glands as described by Marg et al (1992). After purification on a Sephadex G-50 column, a part of the main adrenodoxin fraction intended for fluorescence measurements was subjected to an additional chromatographic step on a HPLC Mono-Q 10/10 column (Pharmacia) and the adrenodoxin peak was eluted with a linear salt gradient (from 0.2 M to 0.4 M KC1 in 20 mM Tris/HCl, pH 7.4).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Mutational Effects on the Spectroscopic Properties and Biological Activities of Oxidized Bovine Adrenodoxin, and Their Structural Implications

Beckert¹,

Schrauber²,

Bernhardt³

et al. 1995

Eur J Biochem

View full text Add to dashboard Cite

Of the aromatic 'H-NMR signals of oxidized bovine adrenodoxin only those of His56 showed intrinsic chemical shift changes upon replacement of Tyr82 by Ser or Leu, that must arise from a loss of a throughspace ring-current effect of the tyrosine ring in these mutants. Thus, of the three His residues contained in adrenodoxin, His56 is closest to Tyr82, and hence to the highly acidic determinant region of adrenodoxin that is the interaction site for adrenodoxin reductase and P-450. The strong dependence of the fluorescence intensity of Tyr82 on the residue in position 56 supported this observation.As a consequence of this, the effects of replacement of His56 by Gln or Thr on cytochrome c reduction and cytochromes P-45OlI, (CYPllB1)-dependent and P-450,,, (CYP1 lA1)-dependent substrate conversions were studied. No influence on V,,, values was observed for all reactions mediated by the mutants, implying His56 does not play a decisive role in the intramolecular or intermolecular electron transfer. In contrast, the K, values were increased, as was the K, value for binding of CYPl 1Al to the IH56Tladreno-doxin.The secondary structure deduced from further NMR data of adrenodoxin was compared with that of other ferredoxins. Tyr82 is in a region of the molecule containing no secondary-structure elements. The data for Tyr82 are in keeping with the biological activities and suggests it is in a flexible, solvent-exposed region of the molecule.Keyword,s. Adrenodoxin ; 'H-NMR ; aromatic region ; fluorescence.Adrenodoxin is a member of the ferredoxin family of proteins, that are widely distributed in bacteria, plants and animals. As a rule, the ferredoxins are low-molecular-mass proteins (6000-25 000 Da) that are negatively charged at neutral pH and all contain iron-sulfur clusters as the redox-active group. Bovine adrenodoxin is involved in two electron-transfer systems in the inner mitochondrial membrane of bovine adrenal cortex. Both systems contain NADPH-dependent adrenodoxin reductase, adrenodoxin and the specific cytochrome P-450, cytochrome P-450,,, (CYPllAl) or cytochrome P-45OI,,, (CYPllBl).The three-dimensional structure of adrenodoxin has not yet been elucidated and there are few data on the structural basis of the mechanism of protein-protein interaction among adrenodoxin and its redox partners. The shuttle model (Lambeth et al., 1979;Hanukoglu and Jefcoate, 1980), a ternary complex formation of adrenodoxin reductase, adrenodoxin, and the cytochrome P-450 (Kido and Kimura, 1979), and a model suggesting Correspondence to R. Bemhardt, Max-Delbriick-Centrum fur Molekulare Medizin,

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Mutational Effects on the Spectroscopic Properties and Biological Activities of Oxidized Bovine Adrenodoxin, and Their Structural Implications

Beckert¹,

Schrauber²,

Bernhardt³

et al. 1995

Eur J Biochem

View full text Add to dashboard Cite

show abstract

“…The first crystals of bovine Adx were already obtained 25 years ago [25] but were not used for structural analysis. Bovine Adx has also been crystallized by Marg et al [26], these crystals, however, contained up to 12 protein molecules in the asymmetric unit and were not further analyzed. Here we report the three-dimensional structure of the Adx variant Adx(4-108), where residues Ser1-Ser3 and Asp109-Glu128 have been removed by genetic engineering.…”

Section: Introductionmentioning

confidence: 99%

New aspects of electron transfer revealed by the crystal structure of a truncated bovine adrenodoxin, Adx(4–108)

et al. 1998

View full text Add to dashboard Cite

The crystal structure of Adx(4-108) provides the first detailed description of a vertebrate [2Fe-2S] ferredoxin and serves to explain a large body of biochemical studies in terms of a three-dimensional structure. The structure suggests how a change in the redox state of the [2Fe-2S] cluster may be coupled to a domain motion of the protein. It seems likely that the clearly asymmetric charge distribution on the surface of Adx(4-108) and the resulting strong molecular dipole are involved in electrostatic steering of the interactions with AR and cytochrome P450.

show abstract

“…Hypertension Therapy -Spironolactone or amiloride can be used to correct hypokalemia and treat slight hypertension (not present in this case). If these agents do not prove to be suffi cient or they are not available calcium channel blockers such as nifedipine or verapamil can be used 9 .…”

Section: Discussionmentioning

confidence: 99%