Rubredoxin from the green sulfur bacterium Chlorobium tepidum functions as an electron acceptor for pyruvate ferredoxin oxidoreductase.
Rubredoxin (Rd) from the moderately thermophilic green sulfur bacterium Chlorobium tepidum was found to function as an electron acceptor for pyruvate ferredoxin oxidoreductase (PFOR). This enzyme, which catalyzes the conversion of pyruvate to acetyl-CoA and CO(2), exhibited an absolute dependence upon the presence of Rd. However, Rd was incapable of participating in the pyruvate synthase or CO(2) fixation reaction of C. tepidum PFOR, for which two different reduced ferredoxins are employed as electron donors. These results suggest a specific functional role for Rd in pyruvate oxidation and provide the initial indication that the two important physiological reactions catalyzed by PFOR/pyruvate synthase are dependent on different electron carriers in the cell. The UV-visible spectrum of oxidized Rd, with a monomer molecular weight of 6500, gave a molar absorption coefficient at 492 nm of 6.89 mM(-1) cm(-1) with an A(492)/A(280) ratio of 0.343 and contained one iron atom/molecule. Further spectroscopic studies indicated that the CD spectrum of oxidized C. tepidum Rd exhibited a unique absorption maximum at 385 nm and a shoulder at 420 nm. The EPR spectrum of oxidized Rd also exhibited unusual anisotropic resonances at g = 9.675 and g = 4.322, which is composed of a narrow central feature with broader shoulders to high and low field. The midpoint reduction potential of C. tepidum Rd was determined to be -87 mV, which is the most electronegative value reported for Rd from any source.
Yoon
K S
KS
Department of Microbiology, The Ohio State University, Columbus, Ohio 43210-1292, USA.
Hille
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R
Hemann
C
C
Tabita
F R
FR
D019414:Chlorobi / Q000737:chemistry*; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D004578:Electron Spin Resonance Spectroscopy; D004583:Electrons; D004591:Electrophoresis, Polyacrylamide Gel; D007658:Ketone Oxidoreductases / Q000378:metabolism*; D050743:Pyruvate Synthase; D012416:Rubredoxins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet
D012416:Rubredoxins; D007658:Ketone Oxidoreductases; D050743:Pyruvate Synthase