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. 1997 Jun;179(11):3793–3796. doi: 10.1128/jb.179.11.3793-3796.1997

Maximum activity of recombinant ribulose 1,5-bisphosphate carboxylase/oxygenase of Anabaena sp. strain CA requires the product of the rbcX gene.

L A Li 1, F R Tabita 1
PMCID: PMC179181  PMID: 9171433

Abstract

Filamentous cyanobacteria of the genus Anabaena contain a unique open reading frame, rbcX, which is juxtaposed and cotranscribed with the genes (rbcL and rbcS) encoding form I ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO). Plasmid constructions containing the genes from Anabaena sp. strain CA were prepared, and expression studies in Escherichia coli indicated that the product of the rbcX gene mimicked the ability of chaperonin proteins to facilitate the proper folding of recombinant RubisCO proteins. The purified recombinant Anabaena sp. strain CA RubisCO, much like the RubisCO enzymes from other cyanobacteria, was shown not to undergo inhibition of activity during a time course experiment, and the properties of this chaperoned recombinant protein appear to be consistent with those of the enzyme isolated from the native organism.

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Selected References

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  1. Andrews T. J. Catalysis by cyanobacterial ribulose-bisphosphate carboxylase large subunits in the complete absence of small subunits. J Biol Chem. 1988 Sep 5;263(25):12213–12219. [PubMed] [Google Scholar]
  2. Badger M. R. Kinetic properties of ribulose 1,5-bisphosphate carboxylase/oxygenase from Anabaena variabilis. Arch Biochem Biophys. 1980 Apr 15;201(1):247–254. doi: 10.1016/0003-9861(80)90509-3. [DOI] [PubMed] [Google Scholar]
  3. Fitchen J. H., Knight S., Andersson I., Branden C. I., McIntosh L. Residues in three conserved regions of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase are required for quaternary structure. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5768–5772. doi: 10.1073/pnas.87.15.5768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gatenby A. A. The properties of the large subunit of maize ribulose bisphosphate carboxylase/oxygenase synthesised in Escherichia coli. Eur J Biochem. 1984 Oct 15;144(2):361–366. doi: 10.1111/j.1432-1033.1984.tb08472.x. [DOI] [PubMed] [Google Scholar]
  5. Gibson J. L., Falcone D. L., Tabita F. R. Nucleotide sequence, transcriptional analysis, and expression of genes encoded within the form I CO2 fixation operon of Rhodobacter sphaeroides. J Biol Chem. 1991 Aug 5;266(22):14646–14653. [PubMed] [Google Scholar]
  6. Gibson J. L., Tabita F. R. The molecular regulation of the reductive pentose phosphate pathway in Proteobacteria and Cyanobacteria. Arch Microbiol. 1996 Sep;166(3):141–150. doi: 10.1007/s002030050369. [DOI] [PubMed] [Google Scholar]
  7. Goloubinoff P., Gatenby A. A., Lorimer G. H. GroE heat-shock proteins promote assembly of foreign prokaryotic ribulose bisphosphate carboxylase oligomers in Escherichia coli. Nature. 1989 Jan 5;337(6202):44–47. doi: 10.1038/337044a0. [DOI] [PubMed] [Google Scholar]
  8. Gurevitz M., Somerville C. R., McIntosh L. Pathway of assembly of ribulosebisphosphate carboxylase/oxygenase from Anabaena 7120 expressed in Escherichia coli. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6546–6550. doi: 10.1073/pnas.82.19.6546. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Keen N. T., Tamaki S., Kobayashi D., Trollinger D. Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene. 1988 Oct 15;70(1):191–197. doi: 10.1016/0378-1119(88)90117-5. [DOI] [PubMed] [Google Scholar]
  10. Larimer F. W., Soper T. S. Overproduction of Anabaena 7120 ribulose-bisphosphate carboxylase/oxygenase in Escherichia coli. Gene. 1993 Apr 15;126(1):85–92. doi: 10.1016/0378-1119(93)90593-r. [DOI] [PubMed] [Google Scholar]
  11. Lee B., Berka R. M., Tabita F. R. Mutations in the small subunit of cyanobacterial ribulose-bisphosphate carboxylase/oxygenase that modulate interactions with large subunits. J Biol Chem. 1991 Apr 25;266(12):7417–7422. [PubMed] [Google Scholar]
  12. Lee B., Tabita F. R. Purification of recombinant ribulose-1,5-bisphosphate carboxylase/oxygenase large subunits suitable for reconstitution and assembly of active L8S8 enzyme. Biochemistry. 1990 Oct 9;29(40):9352–9357. doi: 10.1021/bi00492a007. [DOI] [PubMed] [Google Scholar]
  13. Li L. A., Tabita F. R. Transcription control of ribulose bisphosphate carboxylase/oxygenase activase and adjacent genes in Anabaena species. J Bacteriol. 1994 Nov;176(21):6697–6706. doi: 10.1128/jb.176.21.6697-6706.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mizobata T., Akiyama Y., Ito K., Yumoto N., Kawata Y. Effects of the chaperonin GroE on the refolding of tryptophanase from Escherichia coli. Refolding is enhanced in the presence of ADP. J Biol Chem. 1992 Sep 5;267(25):17773–17779. [PubMed] [Google Scholar]
  15. Paul K., Morell M. K., Andrews T. J. Mutations in the small subunit of ribulosebisphosphate carboxylase affect subunit binding and catalysis. Biochemistry. 1991 Oct 15;30(41):10019–10026. doi: 10.1021/bi00105a029. [DOI] [PubMed] [Google Scholar]
  16. Pridmore R. D. New and versatile cloning vectors with kanamycin-resistance marker. Gene. 1987;56(2-3):309–312. doi: 10.1016/0378-1119(87)90149-1. [DOI] [PubMed] [Google Scholar]
  17. Read B. A., Tabita F. R. Amino acid substitutions in the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase that influence catalytic activity of the holoenzyme. Biochemistry. 1992 Jan 21;31(2):519–525. doi: 10.1021/bi00117a031. [DOI] [PubMed] [Google Scholar]
  18. Tabita F. R., Colletti C. Carbon dioxide assimilation in cyanobacteria: regulation of ribulose, 1,5-bisphosphate carboxylase. J Bacteriol. 1979 Nov;140(2):452–458. doi: 10.1128/jb.140.2.452-458.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tabita F. R., Small C. L. Expression and assembly of active cyanobacterial ribulose-1,5-bisphosphate carboxylase/oxygenase in Escherichia coli containing stoichiometric amounts of large and small subunits. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6100–6103. doi: 10.1073/pnas.82.18.6100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Watson G. M., Tabita F. R. Microbial ribulose 1,5-bisphosphate carboxylase/oxygenase: a molecule for phylogenetic and enzymological investigation. FEMS Microbiol Lett. 1997 Jan 1;146(1):13–22. doi: 10.1111/j.1574-6968.1997.tb10165.x. [DOI] [PubMed] [Google Scholar]

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