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. 1977 Jan;129(1):415–421. doi: 10.1128/jb.129.1.415-421.1977

Quaternary structure and oxygenase activity of D-ribulose-1,5-bisphosphate carboxylase from Hydrogenomonas eutropha.

K Purohit, B A McFadden
PMCID: PMC234941  PMID: 401497

Abstract

Electrophoretically homogeneous ribulose-1,5-bisphosphate (RuBP) carboxylase was obtained from autotropically grown Hydrogenomonas eutropha by sedimentation of the 105,000 X g supernatant in a discontinuous sucrose gradient and by ammonium sulfate fractionation followed by another sucrose gradient centrifugation. The molecular weight of the enzyme determined by light scattering was 490,000 +/- 15,000. The enzyme could be dissociated by sodium dodecyl sulfate into three types of subunits, and the molecular weights (+/- 10%) could be measured. There were two species of large subunits, L and L' (molecular weight 56,000 and 52,000, respectively) and one species of small subunits (molecular weight, 15,000). The mole ratio of L to L' was 5:3, and the overall mole ratio of the small to large subunits was 1.08. The simplest quaternary structure of the enzyme is L5L'3S8. The enzyme contained RuBP oxygenase activity as evidenced by the O2-dependent production of phosphoglycolate and 3-phosphoglyceric acid in equimolar quantities from RuBP.

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Selected References

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  1. Akazawa T., Kondo H., Shimazue T., Nishimura M., Sugiyama T. Further studies on ribulose 1,5-diphosphate carboxylase from Chromatium strain D. Biochemistry. 1972 Mar 28;11(7):1298–1303. doi: 10.1021/bi00757a028. [DOI] [PubMed] [Google Scholar]
  2. Andrews T. J., Lorimer G. H., Tolbert N. E. Ribulose diphosphate oxygenase. I. Synthesis of phosphoglycolate by fraction-1 protein of leaves. Biochemistry. 1973 Jan 2;12(1):11–18. doi: 10.1021/bi00725a003. [DOI] [PubMed] [Google Scholar]
  3. Bowes G., Ogren W. L., Hageman R. H. Phosphoglycolate production catalyzed by ribulose diphosphate carboxylase. Biochem Biophys Res Commun. 1971 Nov 5;45(3):716–722. doi: 10.1016/0006-291x(71)90475-x. [DOI] [PubMed] [Google Scholar]
  4. Dunker A. K., Rueckert R. R. Observations on molecular weight determinations on polyacrylamide gel. J Biol Chem. 1969 Sep 25;244(18):5074–5080. [PubMed] [Google Scholar]
  5. Goldthwaite J. J., Bogorad L. A one-step method for the isolation and determination of leaf ribulose-1,5-diphosphate carboxylase. Anal Biochem. 1971 May;41(1):57–66. doi: 10.1016/0003-2697(71)90191-6. [DOI] [PubMed] [Google Scholar]
  6. Hedrick J. L., Smith A. J. Size and charge isomer separation and estimation of molecular weights of proteins by disc gel electrophoresis. Arch Biochem Biophys. 1968 Jul;126(1):155–164. doi: 10.1016/0003-9861(68)90569-9. [DOI] [PubMed] [Google Scholar]
  7. Kuehn G. D., McFadden B. A. Ribulose 1,5-diphosphate carboxylase from Hydrogenomonas eutropha and Hydrogenomonas facilis. I. Purification, metallic ion requirements, inhibition, and kinetic constants. Biochemistry. 1969 Jun;8(6):2394–2402. doi: 10.1021/bi00834a021. [DOI] [PubMed] [Google Scholar]
  8. Kuehn G. D., McFadden B. A. Ribulose 1,5-diphosphate carboxylase from Hydrogenomonas eutropha and Hydrogenomonas facilis. II. Molecular weight, subunits, composition, and sulfhydryl groups. Biochemistry. 1969 Jun;8(6):2403–2408. doi: 10.1021/bi00834a022. [DOI] [PubMed] [Google Scholar]
  9. Kung S. D., Gray J. C., Wildman S. G., Carlson P. S. Polypeptide composition of fraction 1 protein from parasexual hybrid plants in the genus Nicotiana. Science. 1975 Jan 31;187(4174):353–355. doi: 10.1126/science.187.4174.353. [DOI] [PubMed] [Google Scholar]
  10. Kung S. D., Sakano K., Wildman S. G. Multiple peptide composition of the large and small subunits of Nicotiana tabacum fraction I protein ascertained by fingerprinting and electrofocusing. Biochim Biophys Acta. 1974 Sep 13;365(1):138–147. doi: 10.1016/0005-2795(74)90258-x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lord J. M., Brown R. H. Purification and Some Properties of Chlorella fusca Ribulose 1,5-Diphosphate Carboxylase. Plant Physiol. 1975 Feb;55(2):360–364. doi: 10.1104/pp.55.2.360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McFadden B. A. Autotrophic CO2 assimilation and the evolution of ribulose diphosphate carboxylase. Bacteriol Rev. 1973 Sep;37(3):289–319. doi: 10.1128/br.37.3.289-319.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McFadden B. A., Lord J. M., Rowe A., Dilks S. Composition, quaternary structure, and catalytic properties of D-ribulose-1, 5-bisphosphate carboxylase from Euglena gracilis. Eur J Biochem. 1975 May;54(1):195–206. doi: 10.1111/j.1432-1033.1975.tb04129.x. [DOI] [PubMed] [Google Scholar]
  16. McFadden B. A., Tabita F. R. D-ribulose-1, 5-diphosphate carboxylase and the evolution of autotrophy. Biosystems. 1974 Oct;6(2):93–112. doi: 10.1016/0303-2647(74)90002-1. [DOI] [PubMed] [Google Scholar]
  17. McFadden B. A., Tabita F. R., Kuehn G. D. Ribulose-diphosphate carboxylase from the hydrogen bacteria and Rhodospirillum rubrum. Methods Enzymol. 1975;42:461–472. doi: 10.1016/0076-6879(75)42152-8. [DOI] [PubMed] [Google Scholar]
  18. McFadden B. A. The oxygenase activity of ribulose-1,5-bisphosphate carboxylase from Rhodospirillum rubrum. Biochem Biophys Res Commun. 1974 Sep 9;60(1):312–317. doi: 10.1016/0006-291x(74)90206-x. [DOI] [PubMed] [Google Scholar]
  19. Purohit K., McFadden B. A., Cohen A. L. Purification, quaternary structure, composition, and properties of D-ribulose-1,5-bisphosphate carboxylase from Thiobacillus intermedius. J Bacteriol. 1976 Jul;127(1):505–515. doi: 10.1128/jb.127.1.505-515.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Purohit K., McFadden B. A. Heterogeneity of large subunits of ribulose-1,5-bisphosphate carbosylase from Hydrogenomonas eutropha. Biochem Biophys Res Commun. 1976 Aug 23;71(4):1220–1227. doi: 10.1016/0006-291x(76)90784-1. [DOI] [PubMed] [Google Scholar]
  21. Rohrbach M. S., Humphries B. A., Yost F. J., Jr, Rhodes W. G., Boatman S., Hiskey R. G., Harrison J. H. The reaction of 4,4'-bis-dimethylaminodiphenylcarbinol with the sulfhydryl group. A new reagent for sulfhydryl analysis. Anal Biochem. 1973 Mar;52(1):127–142. doi: 10.1016/0003-2697(73)90338-2. [DOI] [PubMed] [Google Scholar]
  22. Rutner A. C. Estimation of the molecular weight of ribulose diphosphate carboxylase sub-units. Biochem Biophys Res Commun. 1970 Jun 5;39(5):923–929. doi: 10.1016/0006-291x(70)90412-2. [DOI] [PubMed] [Google Scholar]
  23. Rutner A. C., Lane M. D. Nonidentical subunits of ribulose diphosphate carboxylase. Biochem Biophys Res Commun. 1967 Aug 23;28(4):531–537. doi: 10.1016/0006-291x(67)90346-4. [DOI] [PubMed] [Google Scholar]
  24. Sugiyama T., Akazawa T. Structure and function of chloroplast proteins. I. Subunit structure of wheat-fraction-I protein. J Biochem. 1967 Oct;62(4):474–482. doi: 10.1093/oxfordjournals.jbchem.a128691. [DOI] [PubMed] [Google Scholar]
  25. Sugiyama T., Ito T., Akazawa T. Subunit structure of ribulose 1,5-diphosphate carboxylase from Chlorella ellipsoidea. Biochemistry. 1971 Aug 31;10(18):3406–3411. doi: 10.1021/bi00794a014. [DOI] [PubMed] [Google Scholar]
  26. Tabita F. R., McFadden B. A. D-ribulose 1,5-diphosphate carboxylase from Rhodospirillum rubrum. II. Quaternary structure, composition, catalytic, and immunological properties. J Biol Chem. 1974 Jun 10;249(11):3459–3464. [PubMed] [Google Scholar]
  27. Tabita F. R., McFadden B. A. Molecular and catalytic properties of ribulose 1,5-bisphosphate carboxylase from the photosynthetic extreme halophile Ectothiorhodospira halophila. J Bacteriol. 1976 Jun;126(3):1271–1277. doi: 10.1128/jb.126.3.1271-1277.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tabita F. R., McFadden B. A. One-step isolation of microbial ribulose-1,5-diphosphate carboxylase. Arch Microbiol. 1974;99(3):231–240. doi: 10.1007/BF00696237. [DOI] [PubMed] [Google Scholar]
  29. Tabita F. R., McFadden B. A., Pfennig N. D-ribulose-1,5-bisphosphate carboxylase in Chlorobium thiosulfatophilum Tassajara. Biochim Biophys Acta. 1974 Mar 21;341(1):187–194. doi: 10.1016/0005-2744(74)90079-5. [DOI] [PubMed] [Google Scholar]
  30. Tabita F. R., Stevens S. E., Jr, Gibson J. L. Carbon dioxide assimilation in blue-green algae: initial studies on the structure of ribulose 1,5-bisphosphate carboxylase. J Bacteriol. 1976 Feb;125(2):531–539. doi: 10.1128/jb.125.2.531-539.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tabita R. F., Stevens S. E., Jr, Quijano R. D-ribulose 1, 5-diphosphate carboxylase from blue-green algae. Biochem Biophys Res Commun. 1974 Nov 6;61(1):45–52. doi: 10.1016/0006-291x(74)90531-2. [DOI] [PubMed] [Google Scholar]
  32. Takabe T., Akazawa T. Oxidative formation of phosphoglycolate from ribulose-1,5-diphosphate catalysed by Chromatium ribulose-1,5-diphosphate carboxylase. Biochem Biophys Res Commun. 1973 Aug 21;53(4):1173–1179. doi: 10.1016/0006-291x(73)90588-3. [DOI] [PubMed] [Google Scholar]
  33. Takabe T., Nishimura M., Akazawa T. Presence of two subunit types in ribulose-1,5-bisphosphate carboxylase from blue-green algae. Biochem Biophys Res Commun. 1976 Jan 26;68(2):537–544. doi: 10.1016/0006-291x(76)91179-7. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Weber K., Pringle J. R., Osborn M. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol. 1972;26:3–27. doi: 10.1016/s0076-6879(72)26003-7. [DOI] [PubMed] [Google Scholar]

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