A RHIZOBIUM-MELILOTI FERREDOXIN (FDXN) PURIFIED FROM ESCHERICHIA-COLI DONATES ELECTRONS TO RHODOBACTER-CAPSULATUS NITROGENASE

RIEDEL KU, JOUANNEAU Y, MASEPOHL B, Pühler A, KLIPP W (1995)
EUROPEAN JOURNAL OF BIOCHEMISTRY 231(3): 742-746.

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Abstract
The fdxN gene from Rhizobium meliloti encoding a bacterial-type ferredoxin (FdxN) was expressed in Escherichia coil under the control of the lac promoter. The fdxN gene product was purified under anaerobic conditions by ion-exchange chromatography and gel-filtration steps using an antiserum raised against an FdxN-LacZ fusion protein as a detection system. The purified ferredoxin was shown to be identical to the predicted R. meliloti FdxN protein in its amino acid composition and N-terminal amino acid sequence. Chemical determination of the iron content revealed 8.6+/-0.6 mol Fe/mol FdxN. The ultraviolet/visible absorption spectrum of the FdxN protein in the oxidized form exhibited maxima at 284 nm and 378 nm, with an A(378)/A(284) ratio of 0.7. EPR spectroscopy revealed a rhombic signal when FdxN was partially reduced, and a broad signal indicative of spin-spin interaction when fully reduced, suggesting the presence of two Fe-S clusters/ferredoxin polypeptide. Our data suggest that FdxN contains two [4Fe-4S] clusters. Purified FdxN was able to mediate electron transport between illuminated chloroplasts and Rhodobacter capsulatus nitrogenase in vitro.
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RIEDEL KU, JOUANNEAU Y, MASEPOHL B, Pühler A, KLIPP W. A RHIZOBIUM-MELILOTI FERREDOXIN (FDXN) PURIFIED FROM ESCHERICHIA-COLI DONATES ELECTRONS TO RHODOBACTER-CAPSULATUS NITROGENASE. EUROPEAN JOURNAL OF BIOCHEMISTRY. 1995;231(3):742-746.
RIEDEL, K. U., JOUANNEAU, Y., MASEPOHL, B., Pühler, A., & KLIPP, W. (1995). A RHIZOBIUM-MELILOTI FERREDOXIN (FDXN) PURIFIED FROM ESCHERICHIA-COLI DONATES ELECTRONS TO RHODOBACTER-CAPSULATUS NITROGENASE. EUROPEAN JOURNAL OF BIOCHEMISTRY, 231(3), 742-746.
RIEDEL, K. U., JOUANNEAU, Y., MASEPOHL, B., Pühler, A., and KLIPP, W. (1995). A RHIZOBIUM-MELILOTI FERREDOXIN (FDXN) PURIFIED FROM ESCHERICHIA-COLI DONATES ELECTRONS TO RHODOBACTER-CAPSULATUS NITROGENASE. EUROPEAN JOURNAL OF BIOCHEMISTRY 231, 742-746.
RIEDEL, K.U., et al., 1995. A RHIZOBIUM-MELILOTI FERREDOXIN (FDXN) PURIFIED FROM ESCHERICHIA-COLI DONATES ELECTRONS TO RHODOBACTER-CAPSULATUS NITROGENASE. EUROPEAN JOURNAL OF BIOCHEMISTRY, 231(3), p 742-746.
K.U. RIEDEL, et al., “A RHIZOBIUM-MELILOTI FERREDOXIN (FDXN) PURIFIED FROM ESCHERICHIA-COLI DONATES ELECTRONS TO RHODOBACTER-CAPSULATUS NITROGENASE”, EUROPEAN JOURNAL OF BIOCHEMISTRY, vol. 231, 1995, pp. 742-746.
RIEDEL, K.U., JOUANNEAU, Y., MASEPOHL, B., Pühler, A., KLIPP, W.: A RHIZOBIUM-MELILOTI FERREDOXIN (FDXN) PURIFIED FROM ESCHERICHIA-COLI DONATES ELECTRONS TO RHODOBACTER-CAPSULATUS NITROGENASE. EUROPEAN JOURNAL OF BIOCHEMISTRY. 231, 742-746 (1995).
RIEDEL, KU, JOUANNEAU, Y, MASEPOHL, B, Pühler, Alfred, and KLIPP, W. “A RHIZOBIUM-MELILOTI FERREDOXIN (FDXN) PURIFIED FROM ESCHERICHIA-COLI DONATES ELECTRONS TO RHODOBACTER-CAPSULATUS NITROGENASE”. EUROPEAN JOURNAL OF BIOCHEMISTRY 231.3 (1995): 742-746.
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