Skip to main content
SpringerLink
Log in

Mutations in a sequence near the N-terminus of the small subunit alter the CO2/O2specificity factor for ribulose bisphosphate carboxylase/oxygenase

  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

Seven unique substitutions have been introduced by site-directed mutagenesis into the first conserved region of the small subunit of ribulose bisphosphate carboxylase/oxygenase from Anacystis nidulans 6301. After expression of each large, altered-small subunit gene tandem in Escherichia coli, two substitutions in the small subunit tyr17→asp17 (Y17D) and arg10→gly10 (R10G) result in little or no carboxylase activity. For the latter substitution, no L8S8enzyme complex could be detected suggesting that this mutation prevents assembly. Mutant enzymes containing the following substitutions R11G, T14A, S16A, Y17D and P19A have CO2/O2specificity factors (τ values) of 40, 35, 18, 39 and 44, respectively, compared with that of 44 for wild-type recombinant enzyme whereas P20A has full carboxylase activity and a τ value of 55.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Andersen K and Caton J (1987) Sequence analysis of the Alcaligenes eutrophuschromosomally encoded ribulose bisphosphate carboxylase large and small subunit genes and their gene products. J Bacteriol 169: 4547-4558

    PubMed  Google Scholar 

  • Andrews TJ (1988) Catalysis by cyanobacterial ribulose-bisphosphate carboxylase large subunits in the complete absence of small subunits. J Biol Chem 263: 12213-12219

    PubMed  Google Scholar 

  • Boros I, Posfai G and Venetianer P (1984) High-copy-number derivatives of the plasmid cloning vector pBR322. Gene 30: 257-260

    PubMed  Google Scholar 

  • Burke DT and Olson MV (1986) Oligodeoxynucleotide-directed mutagenesis of Escherichia coliand yeast by simple cotransformation of the primer and template. DNA 4: 325-332

    Google Scholar 

  • Carter P, Bedouelle H and Winter G (1985) Improved oligonucleotide site-directed mutagenesis using M13 vectors. Nucleic Acids Res 13: 4431-4443

    PubMed  Google Scholar 

  • Fitchen JH, Knight S, Andersson I, Brändèn CI and McIntosh L (1990) 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 USA 15: 5768-5772

    Google Scholar 

  • Hartman FC and Harpel MR (1994) Structure, function, regulation, and assembly of D-ribulose-1,5-bisphosphate carboxylase/oxygenase. Annu Rev Biochem 63: 197-234

    Article  PubMed  Google Scholar 

  • Jordan DB and Ogren WL Species variation in kinetic properties of ribulose 1,5-bisphosphate carboxylase/oxygenase. Arch Biochem Biophys 1983 7: 425-433

    Google Scholar 

  • Knight S, Andersson I and Brändèn CI (1990) Crystallographic analysis of ribulose 1,5-bisphosphate carboxylase from spinach at 2.4 Å resolution. Interactions and active site. J Mol Biol 215: 113-160

    PubMed  Google Scholar 

  • Kostov RV and McFadden BA (1995) A sensitive simultaneous analysis of ribulose-1,5-bisphosphate carboxylase/ oxygenase efficiencies: Graphical determination of the CO2/O2specificity factor. Photosynth Res 43: 57-66

    Google Scholar 

  • Lee B, Berka RM and Tabita FR (1991) Mutations in the small subunit of cyanobacterial ribulose-bisphosphate carboxylase/oxygenase that modulate interactions with large subunits. J Biol Chem 266: 7417-22

    PubMed  Google Scholar 

  • Lee GJ and McFadden BA (1992) Serine-376 contributes to the binding of substrate by ribulose-bisphosphate carboxylase/oxygenase from Anacystis nidulans. Biochemistry 31: 2304-2308

    PubMed  Google Scholar 

  • Lee GJ, Kostov RV and McFadden BA (1993) A facile method to determine the CO2/O2specificity factor for ribulose-bisphosphate carboxylase/ oxygenase. Photosynth Res 37: 81-86

    Google Scholar 

  • McFadden BA, Tabita FR and Kuehn GD (1975) Ribulose diphosphate carboxylase from the hydrogen bacteria and Rhodospirillum rubrum. Methods Enzymol 42: 461-472

    PubMed  Google Scholar 

  • McFadden BA and Small CL (1988) Cloning, expression and directed mutagenesis of the genes for ribulose bisphosphate carboxylase/oxygenase. Photosynth Res 18: 245-260

    Google Scholar 

  • McFadden, B A, Torres-Ruiz, J A, and Franceschi V R (1989) Localization of ribulose bisphosphate carboxylase/oxygenase and its putative binding protein in the cell envelope of Chromatium vinosum. Planta 178: 297-302

    Google Scholar 

  • McFadden BA (1980) A perspective of ribulose bisphosphate carboxylase/oxygenase, the key catalyst in photosynthesis and photorespiration. Acc Chem Res 13: 394-399

    Google Scholar 

  • Newman J and Gutteridge S (1993) The x-ray structure of Synechococcusof ribulose bisphosphate carboxylase/oxygenase activated quaternary complex between enzyme at 2.2 Å resolution. J Biol Chem 268: 25876-25886

    PubMed  Google Scholar 

  • Nierzwicki Bauer SA, Curtis SE and Haselkorn R (1984) Cotranscription of genes encoding the small and large subunits of ribulose-1,5-bisphosphate carboxylase in the cyanobacterium Anabaena7120. Proc Natl Acad Sci USA 81: 5961-5965

    PubMed  Google Scholar 

  • Norrander J, Kempe T and Messing J (1983) Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene 26: 101-106

    Article  PubMed  Google Scholar 

  • Paul K, Morell MK and Andrews TJ (1991) Mutations in the small subunit of ribulose bisphosphate carboxylase affect subunit binding and catalysis. Biochemistry 30: 10019-10026

    PubMed  Google Scholar 

  • Richardson DC and Richardson JS (1994) Kinemages-simple macromolecular graphics for interactive teaching and publication. Trends Biochem Sci 19: 135-138

    PubMed  Google Scholar 

  • Robison PD, Martin MN and Tabita FR (1979) Differential effects of metal ions on Rhodospirillum rubrumribulose bisphosphate carboxylase/oxygenase and stoichiometric incorporation of HCO3−into a cobalt (III) enzyme complex. Biochemistry 18: 4453-4458

    PubMed  Google Scholar 

  • Sanger F, Nicklen S and Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463-5467

    PubMed  Google Scholar 

  • Shinozaki K and Sugiura M (1983) The gene for the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase is located close to the gene for the large subunit in the cyanobacterium Anacystis nidulans6301. Nucleic Acids Res 11: 6957-6964

    PubMed  Google Scholar 

  • Tabita FR and Small CL (1985) Expression and assembly of active cyanobacterial ribulose-1,5-bisphosphate carboxylase/oxygenase in Escherichia colicontaining stoichiometric amounts of large and small subunits. Proc Natl Acad Sci USA 82: 6100-6103

    PubMed  Google Scholar 

  • Tabita F R (1988) Molecular and cellular regulation of autotrophic carbon dioxide fixation in microorganisms. Microbiol Rev 52: 155-189

    Google Scholar 

  • Torres-Ruiz JA and McFadden BA (1988) A homolog of ribulose bisphosphate carboxylase/oxygenase-binding protein in Chromatium vinosum. Arch Biochem Biophys 261: 196-204

    PubMed  Google Scholar 

  • Torres-Ruiz J and McFadden BA (1985) Isolation of L8and L8S8forms of ribulose bisphosphate carboxylase/oxygenase from Chromatium vinosum. Arch Microbiol 142: 55-60

    Article  PubMed  Google Scholar 

  • Vieira J and Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19: 259-68

    Article  PubMed  Google Scholar 

  • Voordouw G, De Vries PA, Van den Berg WA and De Clerck EP (1987) Site-directed mutagenesis of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase from Anacystis nidulans. Eur J Biochem 163: 591-598

    PubMed  Google Scholar 

  • Zoller MJ and Smith M (1983) Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol 100: 468-500

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry and Biophysics, Washington State University, Pullman, WA, 99164-4660, USA

    Roumen V. Kostov, Christopher L. Small & Bruce A. McFadden

Authors
  1. Roumen V. Kostov
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christopher L. Small
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bruce A. McFadden
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kostov, R.V., Small, C.L. & McFadden, B.A. Mutations in a sequence near the N-terminus of the small subunit alter the CO2/O2specificity factor for ribulose bisphosphate carboxylase/oxygenase. Photosynthesis Research 54, 127–134 (1997). https://doi.org/10.1023/A:1005967106993

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1005967106993

Search

Navigation