Preferences help
enabled [disable] Abstract
Number of results
2008 | 55 | 4 | 777-785
Article title

Heterologous expression and initial characterization of recombinant RbcX protein from Thermosynechococcus elongatus BP-1 and the role of RbcX in RuBisCO assembly

Title variants
Languages of publication
In the cyanobacterial RuBisCO operon from Thermosynechococcus elongatus the rbcX gene is juxtaposed and cotranscribed with the rbcL and rbcS genes which encode large and small RuBisCO subunits, respectively. It has been suggested that the rbcX position is not random and that the RbcX protein could be a chaperone for RuBisCO. In this study, the RbcX protein from T. elongatus was overexpressed, purified and preliminary functional studies were conducted. The recombinant protein purified from Escherichia coli extracts was predominantly present in a soluble fraction in a dimeric form. Coexpression experiments have demonstrated that RbcX can mediate RbcL dimer (L2) formation, and that it is essential for the L8 core complex assembly. This is the first characterization of the RbcX protein from a thermophilic organism.
Physical description
  • Department of Biophysics, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland
  • Department of Biophysics, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland
  • Department of Biophysics, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland
  • Department of Biophysics, Faculty of Biotechnology, University of Wroclaw, Wrocław, Poland
  • Andersson I, Taylor TC (2003) Structural framework for catalysis and regulation in ribulose-1,5-bisphosphate carboxylase/oxygenase. Arch Biochem Biophys 414: 130-140.
  • Bowes G, Ogren WL, Hageman RH (1971) Phosphoglycolate production catalysed by ribulose diphosphate carboxylase. Biochem Biophys Res Commun 45: 716-722.
  • Brinker A, Pfeifer G, Kerner MJ, Naylor DJ, Hartl FU, Hayer-Hartl M (2001) Dual function of protein confinement in chaperonin-assisted protein folding. Cell 107: 223-233.
  • Cleland WW, Andrews TJ, Gutteridge S, Hartman FC, Lorimer GH (1998) Mechanism of RuBisCO: The carbamate as general base. Chem Rev 98: 549-562.
  • Cohen I, Sapir Y, Shapira M (2006) A conserved mechanism controls translation of RuBisCO large subunit in different photosynthetic organisms. Plant Physiol 141: 1089-1097.
  • Ellis RJ (1979) The most abundant protein in the world. Trends Biochem Sci 4: 241-244.
  • Emlyn-Jones D, Woodger FJ, Price GD, Whitney SM (2006) RbcX can function as a RuBisCO-chaperonin, but is not essential in Synechococcus PCC7942. Plant Cell Physiol 47: 1630-1640.
  • Goloubinoff P, Gatenby AA, Lorimer GH (1989a) GroE heat-shock proteins promote assembly of foreign prokaryotic ribulose bisphosphate carboxylase oligomers in Escherichia coli. Nature 337: 44-47.
  • Goloubinoff P, Christeller JT, Gatenby AA, Lorimer GH (1989b) Reconstitution of active dimeric ribulose bisphosphate carboxylase from unfolded state depends on two chaperonin proteins and MgATP. Nature 342: 884-889.
  • Gubernator B, Bartoszewski R, Króliczewski J, Wildner G, Szczepaniak A (2008) Ribulose-1,5-bisphosphate carboxylase/oxygenase from thermophilic cyanobacterium Thermosynechococcus elongatus. Photosynth Res 95: 101-109.
  • Kitano K, Maeda N, Fukui T, Atomi H, Imanaka T, Miki K (2001) Crystal structure of a novel-type archaeal RuBisCO with pentagonal symmetry. Structure 9: 473-481.
  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.
  • Larimer FW, Soper TS (1993) Overproduction of Anabaena 7120 ribulose-bisphosphate carboxylase/oxygenase in Escherichia coli. Gene 126: 85-92.
  • Li L-A, Tabita FR (1997) Maximum activity of recombinant ribulose-1,5-bisphosphate caerboxylase/oxygenase of Anabaena sp. strain CA requires the product of the rbcX gene. J Bacteriol 179: 3793-3796.
  • McKay RML, Gibbs SP, Espie GS (1993) Effect of dissolved inorganic carbon on the expression of carboxysomes, localization of RuBisCO and the mode of inorganic carbon transport in cells of the Synechococcus UTEX 625. Arch Microbiol 199: 21-29.
  • Nakamura Y, Kaneko T, Sato S, Ikeuchi M, Katoh H, Sasamoto S, Watanabe A, Iriguchi M, Kawashima K, Kimura T, Kishida Y, Kiyokawa C, Kohara M, Matsumoto M, Matsuno A, Nakazaki N, Shimpo S, Sugimoto M, Takeuchi C, Yamada M, Tabata S (2002) Complete genome structure of the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1. DNA Res 9: 123-130.
  • Onizuka T, Endo S, Akiyama H, Kanai S, Hirano M, Yokoya A, Tanaka S, Miyasaka H (2004) The rbcX gene product promotes the production and assembly of ribulose-1,5-bisphosphate carboxylase/oxygenase of Synechococcus sp. PCC7002 in Escherichia coli. Plant Cell Physiol 45: 1390-1395.
  • Price GD, Howitt SM, Harrison K, Badger MR (1993) Analysis of a genomic DNA region from the cyanobacterium Synechococcus sp. strain PCC7942 involved in carboxysome assembly and function. J Bacteriol 175: 2871-2879.
  • Rice SC, Pon NG (1978) Direct spectrophotometric observation of ribulose-1,5-bisphosphate carboxylase activity. Anal Biochem 87: 39-48.
  • Rippka R (1988) Isolation and purification of cyanobacteria. Methods Enzymol 167: 3-27.
  • Saschenbrecker S, Bracher A, Rao KV, Rao BV, Hartl FU, Hayer-Hartl M (2007) Structure and function of rbcX, an assembly chaperone for hexadecameric RuBisCO. Cell 129: 1189-1200.
  • Szczepaniak A, Cramer WA (1990) Thylakoid membrane protein topography-location of the termini of the chloroplast cytochrome b6 on the stromal side of the membrane. J Biol Chem 265: 17720-17726.
  • Tabita FR (1999) Microbial ribulose 1,5-bisphosphate carboxylase oxygenase: A different perspective. Photosynth Res 60: 1-28.
  • Whitman W, Tabita FR (1976) Inhibition of d-ribulose 1,5-bisphosphate carboxylase by pyridoxal 5'-phosphate. Biochem Biophys Res Commun 71: 1034-1039.
  • Whitmarsh J, Govindjee (1999) The photosynthetic process. In Photobiology: Photosynthesis and Photomorphogenesis. Singhal GS, Renger G, Sopory SK, Irrgang K-D, Govindjee, eds, pp 11-51. Narosa Publishers/Kluwer Academic Publishers, Dordrecht.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.