Isolation and kinetic characterisation of hydrophobically distinct populations of form I Rubisco

Plant Methods

Laura M C Barter²  Rudiger Woscholski²  Simon Kretschmer¹  Lok Hang Mak¹  M Salman Butt²  Priya Patel¹  Guangyuan Zhao¹  Kerry O’Donnelly² 

[1] Department of Chemistry, Imperial College, South Kensington Campus, Exhibition Road, London SW7 2AZ, UK;Institute of Chemical Biology, Department of Chemistry, Imperial College, Flowers Building, South Kensington Campus, Exhibition Road, London SW7 2AZ, UK

关键词: Spinacia oleracea;    Rubisco;    Purification;    Hydrophobic interaction chromatography;    Brassica oleracea;   

Others  :  802487
DOI  :  10.1186/1746-4811-10-17

 received in 2014-01-10, accepted in 2014-05-28,  发布年份 2014

【 摘 要 】

Background

Rubisco (Ribulose-1,5-bisphosphate carboxylase/oxygenase) is a Calvin Cycle enzyme involved in CO₂ assimilation. It is thought to be a major cause of photosynthetic inefficiency, suffering from both a slow catalytic rate and lack of specificity due to a competing reaction with oxygen. Revealing and understanding the engineering rules that dictate Rubisco’s activity could have a significant impact on photosynthetic efficiency and crop yield.

Results

This paper describes the purification and characterisation of a number of hydrophobically distinct populations of Rubisco from both Spinacia oleracea and Brassica oleracea extracts. The populations were obtained using a novel and rapid purification protocol that employs hydrophobic interaction chromatography (HIC) as a form I Rubisco enrichment procedure, resulting in distinct Rubisco populations of expected enzymatic activities, high purities and integrity.

Conclusions

We demonstrate here that HIC can be employed to isolate form I Rubisco with purities and activities comparable to those obtained via ion exchange chromatography (IEC). Interestingly, and in contrast to other published purification methods, HIC resulted in the isolation of a number of hydrophobically distinct Rubisco populations. Our findings reveal a so far unaccounted diversity in the hydrophobic properties within form 1 Rubisco. By employing HIC to isolate and characterise Spinacia oleracea and Brassica oleracea, we show that the presence of these distinct Rubisco populations is not species specific, and we report for the first time the kinetic properties of Rubisco from Brassica oleracea extracts. These observations may aid future studies concerning Rubisco’s structural and functional properties.

【 授权许可】

   
2014 O’Donnelly et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Spreitzer R, Salvucci M: Rubisco: structure, regulatory interactions, and possibilities for a better enzyme. Annu Rev Plant Biol 2002, 53:449-475.
• [2]Andersson I: Catalysis and regulation in Rubisco. J Exp Bot 2008, 59(7):1555-1568.
• [3]Behrenfield MJ, Randerson JT, Falkowski P: Primary Production of the Biosphere: Integrating Terrestrial and Oceanic Components. Science 1998, 281(5374):237-240.
• [4]Ellis RJ: Biochemistry: Tackling unintelligent design. Nature 2010, 463(7278):164-165.
• [5]Sage RF, Sage TL, Kocacinar F: Photorespiration and the evolution of C4 photosynthesis. Annu Rev Plant Biol 2012, 63:19-47.
• [6]Zhu XG, Portis AR, Long SP: Would transformation of C3 crop plants with foreign Rubisco increase productivity? A computational analysis extrapolating from kinetic properties to canopy photosynthesis. Plant Cell Environ 2004, 27(2):155-165.
• [7]Ellis JR: The most abundant protein in the world. Trends Biochem Sci 1979, 4(11):241-244.
• [8]Andersson I, Backlund A: Structure and function of Rubisco. Plant Physiol Biochem 2008, 46(3):275-291.
• [9]Tabita FR, Satagopan S, Hanson T, Kreel N, Scott S: Distinct form I, II, III, and IV Rubisco proteins from the three kingdoms of life provide clues about Rubisco evolution and structure/function relationships. J Exp Bot 2008, 59(7):1515-1524.
• [10]Chapman MS, Cascio D, Eisenberg D, Suh SW, Smith WW: Sliding-layer conformational change limited by the quaternary structure of plant RuBisCO. Nature 1987, 329(6137):354-356.
• [11]Andersson I, Knight S, Schneider G, Lindqvist Y, Lundqvist T: Crystal-structure of the active-site of Ribulose-Bisphosphate Carboxylase. Nature 1989, 337(6204):229-234.
• [12]Andersson I, Taylor TC: Structural framework for catalysis and regulation in ribulose-1,5-bisphosphate carboxylase/oxygenase. Arch Biochem Biophys 2003, 414(2):130-140.
• [13]Andrews TJ: Catalysis by cyanobacterial ribulose-bisphosphate carboxylase large subunits in the complete absence of small subunits. J Biol Chem 1988, 263(25):12213-12219.
• [14]Lee B, Tabita FR: Purification of recombinant ribulose-1,5-bisphosphate carboxylase/oxygenase large subunits suitable for reconstitution and assembly of active L8S8 enzyme. Biochemistry 1990, 29(40):9352-9357.
• [15]Whitney SM, Sharwood RE, Orr D, White SJ, Alonso H, Galmés J: Isoleucine 309 acts as a C4 catalytic switch that increases ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) carboxylation rate in Flaveria. Proc Natl Acad Sci 2011, 108(35):14688-14693.
• [16]Ishikawa C, Hatanaka T, Misoo S, Miyake C, Fukayama H: Functional Incorporation of Sorghum Small Subunit Increases the Catalytic Turnover Rate of Rubisco in Transgenic Rice. Plant Physiol 2011, 156(3):1603-1611.
• [17]Spreitzer R: Role of the small subunit in ribulose-1,5-bisphosphate carboxylase/oxygenase. Arch Biochem Biophys 2003, 414(2):141-149.
• [18]Uemura K, Anwaruzzaman , Miyachi S, Yokota A: Ribulose-1,5-bisphosphate carboxylase/oxygenase from thermophilic red algae with a strong specificity for CO2 fixation. Biochem Biophys Res Commun 1997, 233(2):568-71.
• [19]Suarez R, Miro M, Cerda V, Perdomo JA, Galmes J: Automated flow-based anion-exchange method for high-throughput isolation and real-time monitoring of RuBisCO in plant extracts. Talanta 2011, 84(5):1259-66.
• [20]Salvucci ME, Portis AR Jr, Ogren WL: Purification of ribulose-1,5-bisphosphate carboxylase/oxygenase with high specific activity by fast protein liquid chromatography. Anal Biochem 1986, 153(1):97-101.
• [21]Jakob R: 2 Quick methods for isolation of Ribulose-1,5-Bisphosphate Carboxylase Oxygenase. Prep Biochem 1988, 18(3):351-360.
• [22]Whitney SM, Sharwood RE: Linked Rubisco subunits can assemble into functional oligomers without impeding catalytic performance. J Biol Chem 2007, 282(6):3809-18.
• [23]Hemmingsen SM, Ellis RJ: Purification and properties of ribulosebisphosphate carboxylase large subunit binding protein. Plant Physiol 1986, 80(1):269-76.
• [24]Carmo-Silva AE, Barta C, Salvucci ME: Isolation of ribulose-1,5-bisphosphate carboxylase/oxygenase from leaves. Methods in molecular biology (Clifton, NJ) 2011, 684:339-47.
• [25]Whitney SM, Kane HJ, Houtz RL, Sharwood RE: Rubisco oligomers composed of linked small and large subunits assemble in tobacco plastids and have higher affinities for CO2 and O2. Plant Physiol 2009, 149(4):1887-95.
• [26]Rosichan JL, Huffaker RC: Source of Endoproteolytic activity associated with purified Ribulose Bisphosphate Carboxylase. Plant Physiol 1984, 75(1):74-77.
• [27]Kreel NE, Tabita FR: Substitutions at Methionine 295 of Archaeoglobus fulgidus Ribulose-1,5-bisphosphate Carboxylase/Oxygenase Affect Oxygen Binding and CO2/O2 Specificity. J Biol Chem 2007, 282(2):1341-1351.
• [28]McCurry SD, Gee R, Tolbert NE: Ribulose-1,5-bisphosphate carboxylase/oxygenase from spinach, tomato, or tobacco leaves. Methods Enzymol 1982, 90 Pt E(Pt E):515-21.
• [29]Parry MAJ, Andralojc PJ, Parmar S, Keys AJ, Habash D, Paul MJ, Alred R, Quick WP, Servaites JC: Regulation of Rubisco by inhibitors in the light. Plant Cell and Environment 1997, 20(4):528-534.
• [30]Parry MAJ, Keys AJ, Madgwick PJ, Carmo-Silva AE, Andralojc PJ: Rubisco regulation: a role for inhibitors. J Exp Bot 2008, 59(7):1569-1580.
• [31]Sage RF: Variation in the k(cat) of Rubisco in C(3) and C(4) plants and some implications for photosynthetic performance at high and low temperature. J Exp Bot 2002, 53(369):609-20.
• [32]Uemura K, Suzuki Y, Shikanai T, Wadano A, Jensen RG, Chmara W, Yokota A: A Rapid and Sensitive Method for Determination of Relative Specificity of RuBisCO from Various Species by Anion-Exchange Chromatography. Plant Cell Physiol 1996, 37(3):325-331.
• [33]Shen JB, Ogren WL: Alteration of spinach ribulose-1,5-bisphosphate carboxylase/oxygenase activase activities by site-directed mutagenesis. Plant Physiol 1992, 99(3):1201-7.
• [34]Shen JB, Orozco EM Jr, Ogren WL: Expression of the two isoforms of spinach ribulose 1,5-bisphosphate carboxylase activase and essentiality of the conserved lysine in the consensus nucleotide-binding domain. J Biol Chem 1991, 266(14):8963-8.
• [35]Portis AR Jr: Rubisco activase - Rubisco’s catalytic chaperone. Photosynth Res 2003, 75(1):11-27.
• [36]Yeoh H-H, Badger MR, Watson L: Variations in Kinetic Properties of Ribulose-1,5-bisphosphate Carboxylases among Plants. Plant Physiol 1981, 67(6):1151-1155.
• [37]Kelly SM, Price NC: The application of circular dichroism to studies of protein folding and unfolding. Biochim Biophys Acta 1997, 1338(2):161-85.
• [38]Kelly SM, Jess TJ, Price NC: How to study proteins by circular dichroism. Biochim Biophys Acta 2005, 1751(2):119-39.
• [39]Greenfield NJ: Applications of circular dichroism in protein and peptide analysis. TrAC Trends Anal Chem 1999, 18(4):236-244.
• [40]Voordouw G, van der Vies SM, Bouwmeister PP: Dissociation of ribulose-1,5-bisphosphate carboxylase/oxygenase from spinach by urea. Eur J Biochem 1984, 141(2):313-8.
• [41]Li G, Mao H, Ruan X, Xu Q, Gong Y, Zang X, Zhao N: Association of heat-induced conformational change with activity loss of Rubisco. Biochem Biophys Res Commun 2002, 290(3):1128-32.
• [42]Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976, 72:248-54.
• [43]Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970, 227(5259):680-5.
• [44]Sulpice R, Tschoep H, Von Korff M, Büssis D, Usadel B, Höhne M, Witucka-Wall H, Altmann T, Stitt M, Gibon Y: Description and applications of a rapid and sensitive non-radioactive microplate-based assay for maximum and initial activity of D-ribulose-1,5-bisphosphate carboxylase/oxygenase. Plant Cell Environ 2007, 30(9):1163-1175.
• [45]Mak L, Vilar R, Woscholski R: Characterisation of the PTEN inhibitor VO-OHpic. J Chem Biol 2010, 3(4):157-163.