Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Jun;130(3):1084–1090. doi: 10.1128/jb.130.3.1084-1090.1977

A four-iron, four-sulfide ferredoxin with high thermostability from Clostridium thermoaceticum.

S S Yang, L G Ljungdahl, J LeGall
PMCID: PMC235330  PMID: 863852

Abstract

A ferredoxin containing only one [Fe4S4] cluster was purified from Clostridium thermoaceticum. It has a molecular weight of about 7,300, a partial specific volume of 0.67, and an isoelectric point of 3.25. Its absorption spectrum has two maxima at 390 nm (epsilon = 16.8 X 10(3)M-1cm-1) and at 280 nm (epsilon = 24.2 X 10(3)M-1cm-1). The absorption at 390 nm is almost half that of other clostridial ferredoxins, which have two [Fe4S4] clusters, and is similar to other ferredoxins with only one [Fe4S4] cluster. The ferredoxin had high thermal stability and retained over 50% of its activity after treatment at 80 degrees C. It functions in the transfer of electrons from pyruvate to nicotinamide adenine dinucleotide phosphate (NADP), which indicates the presence of pyruvate:ferredoxin oxidoreductase and reduced ferredoxin-NADP reductase in C, thermoaceticum. NADPH is used in the synthesis of acetate from CO2 in this organism.

Full text

PDF
1084

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andreesen J. R., Schaupp A., Neurauter C., Brown A., Ljungdahl L. G. Fermentation of glucose, fructose, and xylose by Clostridium thermoaceticum: effect of metals on growth yield, enzymes, and the synthesis of acetate from CO 2 . J Bacteriol. 1973 May;114(2):743–751. doi: 10.1128/jb.114.2.743-751.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brewer J. M., Ashworth R. B. Disc electrophoresis. J Chem Educ. 1969 Jan;46(1):41–45. doi: 10.1021/ed046p41. [DOI] [PubMed] [Google Scholar]
  3. Devenathan T., Akagi J. M., Hersh R. T., Himes R. H. Ferredoxin from two thermophilic clostridia. J Biol Chem. 1969 Jun 10;244(11):2846–2853. [PubMed] [Google Scholar]
  4. Edelstein S. J., Schachman H. K. The simultaneous determination of partial specific volumes and molecular weights with microgram quantities. J Biol Chem. 1967 Jan 25;242(2):306–311. [PubMed] [Google Scholar]
  5. Gottwald M., Andreesen J. R., LeGall J., Ljungdahl L. G. Presence of cytochrome and menaquinone in Clostridium formicoaceticum and Clostridium thermoaceticum. J Bacteriol. 1975 Apr;122(1):325–328. doi: 10.1128/jb.122.1.325-328.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hase T., Ohmiya N., Matsubara H., Mullinger R. N., Rao K. K., Hall D. O. Amino acid sequence of a four-iron-four-sulphur ferredoxin isolated from Bacillus stearothermophilus. Biochem J. 1976 Oct 1;159(1):55–63. doi: 10.1042/bj1590055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Johnson P. W., Canale-Parola E. Properties of rubredoxin and ferredoxin isolated from spirochetes. Arch Mikrobiol. 1973;89(4):341–353. doi: 10.1007/BF00408901. [DOI] [PubMed] [Google Scholar]
  8. LOVENBERG W., BUCHANAN B. B., RABINOWITZ J. C. STUDIES ON THE CHEMICAL NATURE OF CLOSTRIDIAL FERREDOXIN. J Biol Chem. 1963 Dec;238:3899–3913. [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Li L. F., Ljungdahl L., Wood H. G. Properties of Nicotinamide Adenine Dinucleotide Phosphate-Dependent Formate Dehydrogenase from Clostridium thermoaceticum. J Bacteriol. 1966 Aug;92(2):405–412. doi: 10.1128/jb.92.2.405-412.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ljungdahl L. G., Sherod D. W., Moore M. R., Andreesen J. R. Properties of enzymes from Clostridium thermoaceticum and Clostridium formicoaceticum. Experientia Suppl. 1976;26:237–248. doi: 10.1007/978-3-0348-7675-9_20. [DOI] [PubMed] [Google Scholar]
  12. Lode E. T., Murray C. L., Rabinowitz J. C. Apparent oxidation-reduction potential of Clostridium acidi-urici ferredoxin. Effect of pH, ionic strength, and amino acid replacements. J Biol Chem. 1976 Mar 25;251(6):1683–1687. [PubMed] [Google Scholar]
  13. Malkin R., Rabinowitz J. C. Additional observations on the chemistry of clostridial ferredoxin. Biochemistry. 1966 Apr;5(4):1262–1268. doi: 10.1021/bi00868a020. [DOI] [PubMed] [Google Scholar]
  14. Matsubara H., Sasaki R. M. High recovery of tryptophan from acid hydrolysates of proteins. Biochem Biophys Res Commun. 1969 Apr 29;35(2):175–181. doi: 10.1016/0006-291x(69)90263-0. [DOI] [PubMed] [Google Scholar]
  15. Mitchell E. D., Riquetti P., Loring R. H., Carraway K. L. Quaternary structure of Bacillus subtilis -amylase. Anomalous behavior in sodium dodecyl sulfate. Biochim Biophys Acta. 1973 Jan 25;295(1):314–322. doi: 10.1016/0005-2795(73)90099-8. [DOI] [PubMed] [Google Scholar]
  16. Mullinger R. N., Cammack R., Rao K. K., Hall D. O., Dickson D. P., Johnson C. E., Rush J. D., Simopoulos A. Physicochemical characterization of the four-iron-four-sulphide ferredoxin from Bacillus stearothermophilus. Biochem J. 1975 Oct;151(1):75–83. doi: 10.1042/bj1510075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Perutz M. F., Raidt H. Stereochemical basis of heat stability in bacterial ferredoxins and in haemoglobin A2. Nature. 1975 May 15;255(5505):256–259. doi: 10.1038/255256a0. [DOI] [PubMed] [Google Scholar]
  18. Poston J. M., Kuratomi K., Stadtman E. R. The conversion of carbon dioxide to acetate. I. The use of cobalt-methylcobalamin as a source of methyl groups for the synthesis of acetate by cell-free extracts of Clostridium thermoaceticum. J Biol Chem. 1966 Sep 25;241(18):4209–4216. [PubMed] [Google Scholar]
  19. Poston J. M., Stadtman E. R. The conversion of carbon dioxide to acetate. 3. Demonstration of ferredoxin in the system converting Co-14Ch3-cobalamin to acetate. Biochem Biophys Res Commun. 1967 Mar 9;26(5):550–555. doi: 10.1016/0006-291x(67)90100-3. [DOI] [PubMed] [Google Scholar]
  20. Que L., Jr, Holm R. H., Mortenson L. E. Letter: Extrusion of Fe2S2 and Fe4S4 cores from the active sites of ferredoxin proteins. J Am Chem Soc. 1975 Jan 22;97(2):463–464. doi: 10.1021/ja00835a064. [DOI] [PubMed] [Google Scholar]
  21. Rabinowitz J. Preparation and properties of clostridial ferredoxins. Methods Enzymol. 1972;24:431–446. doi: 10.1016/0076-6879(72)24089-7. [DOI] [PubMed] [Google Scholar]
  22. SIEGEL L. M. A DIRECT MICRODETERMINATION FOR SULFIDE. Anal Biochem. 1965 Apr;11:126–132. doi: 10.1016/0003-2697(65)90051-5. [DOI] [PubMed] [Google Scholar]
  23. Schulman M., Ghambeer R. K., Ljungdahl L. G., Wood H. G. Total synthesis of acetate from CO2. VII. Evidence with Clostridium thermoaceticum that the carboxyl of acetate is derived from the carboxyl of pyruvate by transcarboxylation and not by fixation of CO2. J Biol Chem. 1973 Sep 25;248(18):6255–6261. [PubMed] [Google Scholar]
  24. Strombaugh N. A., Burris R. H., Orme-Johnson W. H. Ferredoxins from Bacillus polymyxa. Low potential iron-sulfur proteins which appear to contain single four iron, four sulfur centers accepting a single electron on reduction. J Biol Chem. 1973 Nov 25;248(22):7951–7956. [PubMed] [Google Scholar]
  25. Thauer R. K. CO(2)-reduction to formate by NADPH. The initial step in the total synthesis of acetate from CO(2) in Clostridium thermoaceticum. FEBS Lett. 1972 Oct 15;27(1):111–115. doi: 10.1016/0014-5793(72)80421-6. [DOI] [PubMed] [Google Scholar]
  26. Travis J., Newman D. J., LeGall J., Peck H. D., Jr The amino acid sequence of ferredoxin from the sulfate reducing bacterium, Desulfovibrio gigas. Biochem Biophys Res Commun. 1971 Oct 15;45(2):452–458. doi: 10.1016/0006-291x(71)90840-0. [DOI] [PubMed] [Google Scholar]
  27. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  28. Yoch D. C., Arnon D. I., Sweeney W. V. Characterization of two soluble ferredoxins as distinct from bound iron-sulfur proteins in the photosynthetic bacterium Rhodospirillum rubrum. J Biol Chem. 1975 Nov 10;250(21):8330–8336. [PubMed] [Google Scholar]
  29. Yoch D. C. Purification and properties of two ferredoxins from the nitrogen-fixing bacterium Bacillus polymyxa. Arch Biochem Biophys. 1973 Oct;158(2):633–640. doi: 10.1016/0003-9861(73)90555-9. [DOI] [PubMed] [Google Scholar]
  30. Yoch D. C., Valentine R. C. Four-iron (sulfide) ferredoxin from Bacillus polymyxa. J Bacteriol. 1972 Jun;110(3):1211–1213. doi: 10.1128/jb.110.3.1211-1213.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zubieta J. A., Mason R., Postgate J. R. A four-iron ferredoxin from Desulfovibrio desulfuricans. Biochem J. 1973 Aug;133(4):851–854. doi: 10.1042/bj1330851. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES