title: Structural rearrangements of the two domains of Azotobacter vinelandii rhodanese upon sulfane sulfur release: essential molecular dynamics, 15N NMR relaxation and deuterium exchange on the uniformly labeled protein.
abstract: The Azotobacter vinelandii rhodanese is a 31kDa sulfurtransferase protein that catalyzes the transfer of sulfur atom from thiosulfate to cyanide in the detoxification process from cyanide and is able to insert sulfur atom in the iron-sulfur cluster. A study of the uniformly 15N isotopic labeling by high resolution NMR, before obtaining the backbone sequential assignment, has been carried out. The sulfur loaded and the sulfur discharged forms of the enzyme show very similar HSQC spectra with a good spectral dispersion. Few resonances show changes in chemical shift between the two forms. Relaxation parameters T(1), T(2) and 1H-15N NOE of all amide nitrogen atoms, as well as isotope exchange kinetics, show that the two forms exhibit the same global correlation time and hydrodynamic properties. In parallel, essential dynamics studies show that formation and discharging of catalytic cysteine persulfide group has no significant impact on the overall conformation of the protein. These results, taken together, give a clearcut answer to the question if the catalytic mechanism of the enzyme involves a change in the conformation and/or in the mutual orientation of the two domains. On the contrary these results clearly indicate that upon the catalytic mechanism the two domains of the protein behave as a unique fold.
authors: INFM, Sez. B, University of Rome Tor Vergata, Rome, Italy
authors: Daniel Oscar
authors: DO
authors: Cicero
authors: Sonia
authors: S
authors: Melino
authors: Maria
authors: M
authors: Orsale
authors: Giuseppe
authors: G
authors: Brancato
authors: Andrea
authors: A
authors: Amadei
authors: Fabio
authors: F
authors: Forlani
authors: Silvia
authors: S
authors: Pagani
authors: Maurizio
authors: M
authors: Paci
mesh_terms: D016948:Azotobacter vinelandii / Q000201:enzymology*; D003198:Computer Simulation; D003903:Deuterium; D007553:Isotope Labeling; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009587:Nitrogen Isotopes; D011487:Protein Conformation; D012997:Solvents; D013455:Sulfur / Q000737:chemistry; D013884:Thiosulfate Sulfurtransferase / Q000737:chemistry*
chemical_list: D009587:Nitrogen Isotopes; D012997:Solvents; D013455:Sulfur; D003903:Deuterium; D013884:Thiosulfate Sulfurtransferase