Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1973 Nov;135(3):531–535. doi: 10.1042/bj1350531

Nitrogenase of Klebsiella pneumoniae. Interaction of the component proteins studied by ultracentrifugation

Robert R Eady 1
PMCID: PMC1165855  PMID: 4589392

Abstract

Sedimentation-velocity analyses of mixtures of the component proteins of nitrogenase of Klebsiella pneumoniae at a 1:1 molar ratio, showed a single peak of sedimentation coefficient (12.4S) considerably greater than that obtained for the larger (Fe+Mo-containing) protein centrifuged alone (10.4S). When the ratio exceeded 1:1 (the smaller Fe-containing protein in excess) an additional peak corresponding in sedimentation coefficient (about 4.5S) to free Fe-containing protein appeared. When proteins, which had been inactivated by exposure to air were used, no interaction occurred. Na2S2O4 at 2mm both reversed and prevented interaction between the two proteins; sedimentation coefficients corresponded to those of the proteins when centrifuged alone. These results demonstrate the formation of a complex between the nitrogenase proteins, and, together with data of activity titration curves, are consistent with the formulation of the nitrogenase complex of K. pneumoniae as (Fe-containing protein)–(Fe+Mo-containing protein).

Full text

PDF

Images in this article

533Fig. 1.
on p.533

Selected References

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

  1. Bergersen F. J., Turner G. L. Kinetic studies of nitrogenase from soya-bean root-nodule bacteroids. Biochem J. 1973 Jan;131(1):61–75. doi: 10.1042/bj1310061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Burris R. H. Progress in the biochemistry of nitrogen fixation. Proc R Soc Lond B Biol Sci. 1969 Apr 1;172(1029):339–354. doi: 10.1098/rspb.1969.0025. [DOI] [PubMed] [Google Scholar]
  3. Eady R. R., Smith B. E., Cook K. A., Postgate J. R. Nitrogenase of Klebsiella pneumoniae. Purification and properties of the component proteins. Biochem J. 1972 Jul;128(3):655–675. doi: 10.1042/bj1280655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Huang T. C., Zumft W. G., Mortenson L. E. Structure of the molybdoferredoxin complex from Clostridium pasteurianum and isolation of its subunits. J Bacteriol. 1973 Feb;113(2):884–890. doi: 10.1128/jb.113.2.884-890.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ljones T., Burris R. H. ATP hydrolysis and electron transfer in the nitrogenase reaction with different combinations of the iron protein and the molybdenum-iron protein. Biochim Biophys Acta. 1972 Jul 12;275(1):93–101. doi: 10.1016/0005-2728(72)90027-8. [DOI] [PubMed] [Google Scholar]
  6. Orme-Johnson W. H., Hamilton W. D., Jones T. L., Tso M. Y., Burris R. H., Shah V. K., Brill W. J. Electron paramagnetic resonance of nitrogenase and nitrogenase components from Clostridium pasteurianum W5 and Azotobacter vinelandii OP. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3142–3145. doi: 10.1073/pnas.69.11.3142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Silverstein R., Bulen W. A. Kinetic studies of the nitrogense-catalyzed hydrogen volution and nitrogen reduction reactions. Biochemistry. 1970 Sep 15;9(19):3809–3815. doi: 10.1021/bi00821a021. [DOI] [PubMed] [Google Scholar]
  8. Smith B. E., Lowe D. J., Bray R. C. Nitrogenase of Klebsiella pneumoniae: electron-paramagnetic-resonance studies on the catalytic mechanism. Biochem J. 1972 Nov;130(2):641–643. doi: 10.1042/bj1300641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Smith B. E., Lowe D. J., Bray R. C. Studies by electron paramagnetic resonance on the catalytic mechanism of nitrogenase of Klebsiella pneumoniae. Biochem J. 1973 Oct;135(2):331–341. doi: 10.1042/bj1350331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Thorneley R. N., Eady R. R. Nitrogenase of Klebsiella pneumoniae: evidence for an adenosine triphosphate-induced association of the iron-sulphur protein. Biochem J. 1973 Jun;133(2):405–408. doi: 10.1042/bj1330405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Tso M. Y., Ljones T., Burris R. H. Purification of the nitrogenase proteins from Clostridium pasteurianum. Biochim Biophys Acta. 1972 Jun 23;267(3):600–604. doi: 10.1016/0005-2728(72)90193-4. [DOI] [PubMed] [Google Scholar]
  12. Vandecasteele J. P., Burris R. H. Purification and properties of the constituents of the nitrogenase complex from Clostridium pasteurianum. J Bacteriol. 1970 Mar;101(3):794–801. doi: 10.1128/jb.101.3.794-801.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Yates M. G. Effect of non-haem iron proteins and cytochrome C from Azotobacter upon the activity and oxygen sensitivity of Azobacter nitrogenase. FEBS Lett. 1970 Jun 27;8(5):281–285. doi: 10.1016/0014-5793(70)80287-3. [DOI] [PubMed] [Google Scholar]
  14. Zumft W. G., Cretney W. C., Huang T. C., Mortenson L. E., Palmer G. On the structure and function of nitrogenase from Clostridium pasteurianum W5. Biochem Biophys Res Commun. 1972 Sep 26;48(6):1525–1532. doi: 10.1016/0006-291x(72)90887-x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES