Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1975 Feb;145(2):391–396. doi: 10.1042/bj1450391

Nitrogenase of Klebsiella pneumoniae. A stopped-flow study of magnesium-adenosine triphosphate-induce electron transfer between the compeonent proteins.

R N Thorneley
PMCID: PMC1165229  PMID: 1098654

Abstract

Stopped-flow kinetic data have been obtained for a rapid electron-transfer reaction between the component proteins of nitrogenase from Klebsiella pneumoniae, which was induced by MgATP. Up to three equivalents of the Fe-containing protein were rapidly oxidized by one equivalent of the Fe-Mo-containing protein in a unimolecular reaction, k2 = 2 x 10(2)S-1. Evidence for a tight complex between the component proteins, KD(complex) less than 0.5 muM, which was formed with a rate k1 greater than 1 x 10(7)M-1-S-1, has been obtained. MgATP bound to either the Fe-containing protein or to the two-protein complex with a rate k3 greater than 2.5 x 10(6)M-1-S-1 and with KD(MgATP) = 0.4mM, before the electron-transfer reaction.

Full text

PDF
391

Images in this article

393Fig. 1.
on p.393

Selected References

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

  1. Beinert H. EPR studies on the mechanism of action of succinate dehydrogenase in activated preparations. Biochem Biophys Res Commun. 1974 Jun 4;58(3):564–572. doi: 10.1016/s0006-291x(74)80457-2. [DOI] [PubMed] [Google Scholar]
  2. Bray G. A., Gallagher T. F., Jr Suppression of appetite by bile acids. Lancet. 1968 May 18;1(7551):1066–1067. doi: 10.1016/s0140-6736(68)91415-3. [DOI] [PubMed] [Google Scholar]
  3. Eady R. R. Nitrogenase of Klebsiella pneumoniae. Interaction of the component proteins studied by ultracentrifugation. Biochem J. 1973 Nov;135(3):531–535. doi: 10.1042/bj1350531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Ljones T. Nitrogenase from Clostridium pasteurianum. Changes in optical absorption spectra during electron transfer and effects of ATP, inhibitors and alternative substrates. Biochim Biophys Acta. 1973 Sep 15;321(1):103–113. doi: 10.1016/0005-2744(73)90064-8. [DOI] [PubMed] [Google Scholar]
  6. Mortenson L. E., Zumpft W. G., Palmer G. Electron paramagnetic resonance studies on nitrogenase. 3. Function of magnesium adenosine 5'-triphosphate and adenosine 5'-diphosphate in catalysis by nitrogenase. Biochim Biophys Acta. 1973 Feb 22;292(2):422–435. doi: 10.1016/0005-2728(73)90048-0. [DOI] [PubMed] [Google Scholar]
  7. Nakos G., Mortenson L. Subunit structure of azoferredoxin from Clostridium pasteurianum W5. Biochemistry. 1971 Feb 2;10(3):455–458. doi: 10.1021/bi00779a016. [DOI] [PubMed] [Google Scholar]
  8. Olson J. S., Ballou D. P., Palmer G., Massey V. The mechanism of action of xanthine oxidase. J Biol Chem. 1974 Jul 25;249(14):4363–4382. [PubMed] [Google Scholar]
  9. 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]
  10. Shah V. K., Brill W. J. Nitrogenase. IV. Simple method of purification to homogeneity of nitrogenase components from Azotobacter vinelandii. Biochim Biophys Acta. 1973 May 30;305(2):445–454. doi: 10.1016/0005-2728(73)90190-4. [DOI] [PubMed] [Google Scholar]
  11. Smith B. E., Lang G. Mössbauer spectroscopy of the nitrogenase proteins from Klebsiella pneumoniae. Structural assignments and mechanistic conclusions. Biochem J. 1974 Feb;137(2):169–180. doi: 10.1042/bj1370169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Thorneley R. N. A convenient electrochemical preparation of reduced methyl viologen and a kinetic study of the reaction with oxygen using an anaerobic stopped-flow apparatus. Biochim Biophys Acta. 1974 Mar 26;333(3):487–496. doi: 10.1016/0005-2728(74)90133-9. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Thorneley R. N., Willison K. R. Nitrogenase of Klebsiella pneumoniae. Inhibition of acetylene reduction by magnesium ion explained by the formation of an inactive dimagnesium-adenosine triphophate complex. Biochem J. 1974 Apr;139(1):211–214. doi: 10.1042/bj1390211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tso M. Y., Burris R. H. The binding of ATP and ADP by nitrogenase components from Clostridium pasteurianum. Biochim Biophys Acta. 1973 Jun 6;309(2):263–270. doi: 10.1016/0005-2744(73)90024-7. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Tso M. Y. Some properties of the nitrogenase proteins from Clostridium pasteurianum. Molecular weight, subunit structure, isoelectric point and EPR spectra. Arch Microbiol. 1974;99(1):71–80. doi: 10.1007/BF00696223. [DOI] [PubMed] [Google Scholar]
  20. Walker M., Mortenson L. E. Oxidation reduction properties of nitrogenase from Clostridium pasteurianum W5. Biochem Biophys Res Commun. 1973 Sep 18;54(2):669–676. doi: 10.1016/0006-291x(73)91475-7. [DOI] [PubMed] [Google Scholar]
  21. Zumft W. G., Mortenson L. E., Palmer G. Electron-paramagnetic-resonance studies on nitrogenase. Investigation of the oxidation-reduction behaviour of azoferredoxin and molybdoferredoxin with potentiometric and rapid-freeze techniques. Eur J Biochem. 1974 Aug 1;46(3):525–535. doi: 10.1111/j.1432-1033.1974.tb03646.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES