Conformational change of adrenodoxin induced by reduction of iron-sulfur cluster. Proton nuclear magnetic resonance study.
Bovine adrenodoxin in the reduced form has been measured by one- and two-dimensional 1H NMR spectroscopy. By comparing the spectrum of reduced adrenodoxin with that of the oxidized protein, resonances have been assigned for the aromatic residues. The spin-lattice relaxation time for the resonances due to histidine residues was found to depend on the reduction state of adrenodoxin. The distance from the paramagnetic center is calculated by using the Solomone-Bloembergen equation. The resonances from Tyr-82 and Ala-81 show large chemical shift changes upon reduction of adrenodoxin. The conformational change of adrenodoxin manifested by chemical shift difference between reduced and oxidized forms is found in the sequence around Tyr-82 and Ala-81. Modification with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide at Glu-74, Asp-79, and Asp-86 inhibited the interaction with both adrenodoxin reductase and cytochrome P-450scc (Lambeth, D. J., Geren, L. M., and Millett, F. (1984) J. Biol. Chem. 259, 10025-10029; Geren, L. M., O'Brien, P., Stonehuerner, J., and Millett, F. (1984) J. Biol. Chem. 259, 2155-2160). Thus, the sequence of these amino acids was assigned to the interaction site with the redox partners. The present 1H NMR investigation of adrenodoxin demonstrates that a conformational change upon reduction of the iron-sulfur cluster occurs in the sequence of negatively charged amino acids that is a putative site for interaction with redox partners. This could offer the structural basis of the electron transfer mechanism in which adrenodoxin functions as a mobile electron carrier.
Miura
S
S
Department of Biochemistry, Kagawa Medical School, Japan.
Ichikawa
Y
Y
D000325:Adrenodoxin / Q000737:chemistry*; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002417:Cattle; D007501:Iron / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D010084:Oxidation-Reduction; D011487:Protein Conformation; D011522:Protons; D013455:Sulfur / Q000737:chemistry*; D013696:Temperature
D000596:Amino Acids; D011522:Protons; D000325:Adrenodoxin; D013455:Sulfur; D007501:Iron