Showing papers by "Manuela M. Pereira published in 1999"

Membrane-Bound Electron Transfer Chain of the Thermohalophilic Bacterium Rhodothermus marinus: A Novel Multihemic Cytochrome bc, a New Complex III†

Abstract: A novel multihemic cytochrome bc complex was isolated from the membranes of Rhodothermus marinus. It is a complex with a minimum of three subunits (43, 27, and 18 kDa), containing five low-spin heme centers of the B and C types, in a 1:4 ratio. All the C-type hemes are in the same subunit (27 kDa). Three distinct redox transitions, at 235, 80, and −45 mV, were observed by visible redox titrations. The first involves one B- and one C-type hemes, and in the other two transitions one and two C-type hemes are involved, respectively. Spectroscopic data strongly suggest that the two hemes intervening in the last transition are in van der Waals contact, yielding a split Soret band. Electron paramagnetic resonance spectra of the oxidized complex show resonances of five low-spin ferric heme centers. Upon reduction with ascorbate, all these resonances vanish and a new one attributed to the last pair of hemes appears. A [3Fe-4S]1+/0 center copurifies with this complex, having a high reduction potential of +140 mV. N...

Membrane-bound electron transfer chain of the thermohalophilic bacterium Rhodothermus marinus: characterization of the iron-sulfur centers from the dehydrogenases and investigation of the high-potential iron-sulfur protein function by in vitro reconstitution of the respiratory chain.

Abstract: Rhodothermus marinus, a thermohalophilic bacterium, has a unique electron-transfer chain, containing, besides a cbb3 and a caa3 terminal oxidases, a novel cytochrome bc complex [Pereira, M. M., Carita, J. N., and Teixeira, M. (1999) Biochemistry 38, 1268−1275]. The membrane-bound iron−sulfur centers of this bacterium were studied by electron paramagnetic resonance (EPR) spectroscopy, leading to the identification of its main electron-transfer complexes. The resonances typical for the Rieske-type centers are not detected. Clusters S1 and S3 from succinate dehydrogenase were identified; interestingly, center S3 is shown to be present in two different conformations, with g values at 2.035, 2.009, and 2.001 and at 2.025, 2.002, and 2.000. Upon addition of NADH and dithionite, EPR signals assigned to resonances characteristic of binuclear and tetranuclear clusters develop and are attributed to the iron−sulfur centers of complexes I and II. A high-potential iron−sulfur protein- (HiPIP-) type center previously d...