Abstract
Having a photocatalyzed characteristic, our previous research had proved that nano-anatase TiO2 is closely related to the photosynthesis of spinach. It could not only improve the light absorbance and the transformation from light energy to electron energy and to active chemical energy but also promote carbon dioxide (CO2) assimilation of spinach. However, the mechanism of carbon reaction promoted by nano-anatase TiO2 remains largely unclear. By electrophoresis and Western blot methods, the results of the experiments proved that Rubisco from the nano-anatase TiO2-treated spinach during the extraction procedure of Rubisco was found to consist of Rubisco and a heavier molecular-mass protein (about 1200 kDa) comprising both Rubisco and Rubisco activase. The Rubisco carboxylase activity was 2.67 times that of Rubisco from the control and it could hydrolyze ATP in the same manner as Rubisco activase. The total sulfhydryl groups and available sulfhydryl groups of the Rubisco were 32-SH and 21-SH per mole of enzyme more than those of the Rubisco purified from the control, respectively. The circular dichroism spectra showed that the secondary structure of Rubisco from the nano-anatase TiO2-treated spinach was very different from Rubisco of the control. It suggested that the mechanism of nano-anatase TiO2 activating Rubisco of spinach was that the complex of Rubsico and Rubisco activase was induced in spinach, which promoted Rubsico carboxylation and increased the rate of photosynthetic carbon reaction.
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L. J. Wang, Z. M. Guo, T. J. Li, and M. Li, Biomineralized nanostructured materials and plant silicon nutrition, Prog. Chem. 11, 119–128 (1999) (in Chinese).
R. H. Crabtree, A new type of hydrogen bond, Science 282, 2000–2001 (1998).
L. Zheng, F. S. Hong, S. P. Lu, and C. Liu, Effect of nano-TiO2 on strength of naturally aged seeds and growth of spinach, Biol. Trace Element Res. 104(1), 82–93 (2005).
F. S. Hong, P. Yang, F. Q. Gao, et al., Effect of nano-anatase TiO2 on spectral characterization of photosystem II particles from spinach, Chem. Res. Chin. Univ. 21(2), 196–200 (2005).
F. S. Hong, F. Yang, C. Liu, et al., Influences of nano-TiO2 on the chloroplast ageing of spinach under light, Biol. Trace Element Res. 104(3), 249–260 (2005).
F. S. Hong, J. Zhou, C. Liu, et al., Effect of nano-TiO2 on photochemical reaction of chloroplasts of spinach, Biol. Trace Element Res. 105(3), 269–280 (2005).
J. S. Robert, Questions about the complexity of chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase, Photosynth. Res. 60, 29–42 (1999).
M. A. Parry, P. J. Andralojc, R. A. C. Mitchell, P. J. Madgwick, and A. J. Keys, Manipulation of Rubisco: the amount, activity, function and regulation, J. Exp. Bot. 54(386), 1321–1333 (2003).
A. R. Portis, Jr., Rubisco activase: Rubisco's catalytic chaperone, Photosynth. Res. 75(1), 11–27 (2003).
A. R. Portis, Jr., M. E. Salvucc, and W. L. Ogren, Activation of ribulosebisphosphate carboxylase oxygenase at physiological CO2 and ribulosebisphosphate concentrations by Rubisco activase, Plant Physiol. 82, 967–971 (1986).
A. R. Portis, Jr., The regulation of Rubisco by Rubisco activase, J. Exp. Bot. 46, 1285–1291 (1995).
E. Sánchez de Jiménez, L. Medrano, and E. Mart ble new member of the molecular chaperone family, Biochemistry 34, 2826–2831 (1995).
S. P. Robinson, V. J. Streusand, J. M. Chatifield, et al., Purification and assay of Rubisco activase from leaves, Plant Physiol. 88, 1008–1014 (1988).
Y. Lan and K. A. Mott, Determination of apparent Km values for ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase using the spectrophotometric assay of Rubisco activity, Plant Physiol. 95, 604–609 (1991).
R. H. Tang and L. R. Li, The progress of studies on Rubisco activase, Chin. Bull. Sci. 10(4), 159–166 (1998) (in Chinese).
Y. Han, G. Chen, and Z. Wang, The progress of studies on Rubisco activase, Chin. Bull. Bot. 17(4), 306–311 (2000) (in Chinese).
W. F. Li, Z. Wang, and Y. Han, Purification and activity characteristic of rubisco activase from wheat leaves, Sci. Agric. Sin. 35(8), 929–933 (2002) (in Chinese).
R. J. Spreitzer, Questions about the complexity of chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase, Photosynth. Res. 60, 29–42 (1999).
F. S. Hong, C. Liu, L. Zheng, et al., Formation of complexes of Rubisco-Rubisco activase from La3+, Ce3+ treatment of Spinach, Sci. China S B 48(1), 67–74 (2005).
B. B. Buchanan, W. Gruissem, and R. L. Jones, Biochemistry and Molecular Biology of Plants, Science Press American Society of Plant Physiologists, Beijing, pp. 610–626 (2002).
D. I. Arnon, Copper enzymes in isolated chloroplasts: polyphenol oxidase in Beta vulgaris, Plant Physiol. 24, 1–15 (1949).
W. G. Wang and L. R. Li, A simplified purification method of RuBP carboxylase from spinach leaves, Acta Phytophysiol. Sin. 40(3), 256–262 (1980) (in Chinese).
T. Sugiyama, N. Nakayama, M. Ogawa, T. Akazawa, and T. Oda, Structure and function of chloroplast proteins: effect of ρ-chloromercuribenzoate treatment on the ribulose-1,5-bisphosphate carboxylase/oxygenase activity of spinach leaf fraction protein, Arch. Biochem. Biophys. 125, 98–106 (1968).
O. H. Lowry, N. J. Rosebrough, and A. L. Farr, Protein measurement with the folin phenol reagent, J. Biol. Chem. 193, 265–275 (1951).
F. J. Van de Loo and M. E. Salvucci, Activation of Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) involves Rubisco activase Trp 16, Biochemistry 35, 8143–8148 (1996).
U. K. Laemmli, Cleavage of structural proteins during the assembly of the head of the bacteriophage T4, Nature 277, 680–685 (1970).
E. F. Sambrook and T. Fritsch, eds., Molecular Cloning, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 739–775 (1989).
K. Y. To, M. C. Cheng, L. F. O. Chen, and S. C. G. Chen, Introduction and expression of foreign DNA in isolated spinach chloroplasts by electroporation, Plant J, 10, 737–743 (1996).
A. F. S. A. Habeeb, Reaction of protein sufhydryl group with Ellman's reagent, Methods Enzymol, 227, 457–464 (1972).
A. Perczel, K. Park, and G. D. Fasman, Analysis of the circular dichroism spectrum of proteins using the convex constraint algorithm: A practical guide, Anal. Biochem. 203, 83–89 (1992).
A. F. Neuwald, L. Aravind, J. L. Spouge, and E. V. Koonin, AAA+: a class of chaperone-like ATPases associated with the assembly, operation, and disassembly of protein complexes, Genome Res. 9, 27–43 (1999).
R. J. Spreitzer and M. E. Salvucci, RUBISCO: structure, regulatory interactions, and posibilities for a better enzyme, Annu. Rev. Plant Biol. 53, 449–485 (2002).
Z. L. Zhang and S. Komatsu, Molecular cloning and characterization of cDNAs encoding two isoforms of ribulose-1,5-bisphosphate carboxylase/oxygenase activase in rice (Oryza sativa L.), J. Biochem. 128, 383–389 (2000).
A. Yokota and N. Tsujimoto, Characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase carrying ribulose 1,5-bisphosphate at its regulatory sites and the mechanism of interaction of this form of the enzyme with ribulose-1,5-bisphosphatecarboxylase/oxygenase activase, Eur. J. Biochem. 204, 901–909 (1992).
C. Büchen-Osmond, A. R. Portis, Jr., and T. J. Andrews, Rubisco activase modifies the appearance of Rubisco in the electron microscope, in Research in Photosynthesis, Volume III, IXth International Congress on Photosynthesis, Nagoya, Japan, August 30-September 4, 1992, N. Murata, ed., Kluwer Academic, Dordrecht, pp. 653–656 (1992).
A. R. Portis, Jr., The Rubisco activase-Rubisco system: an ATPase-dependent association that regulates photosynthesis, in Protein-Protein Interactions in Plant Biology, M. T. McManus, W. L. Laing, and A. C. Allen, eds., Sheffield Academic, Sheffield, UK, pp. 30–52 (2001).
M. E. Salvucci, Subunit interactions of Rubisco activase—polyethylene glycol promotes self-association, stimulates ATPase and activation activities, and enhances interactions with Rubisco, Arch. Biochem. Biophys. 298, 688–696 (1992).
Y. G. Miao and L. R. Li, Purification of Rubisco from rice and its properties compared with those of tobacco, Acta Phytophysiol. Sini. 17(2), 183–191 (1991) (in Chinese).
T. Takabe and T. Akazawa, The role of sulfhydrl groups in the ribulose-1,5-bisphosphate carboxylase and oxygenase reaction, Arch. Biochem. Biophys. 169, 686–694 (1975).
R. Chollet and L. L. Anderson, Conformational changes associated with the reversible cold inactivation of ribulose-1,5-bisphosphate carboxylase/oxygenase, Biochem. Biophys. Acta 482, 228–240 (1977).
Y. Tomimatsu and J. W. Donovan, Effect of pH, Mg2+, CO2 and mercurials on the circular dichroism, thermal stability and light scattering of ribulose 1,5-bisphosphate carboxylases from alfalfa, spinach and tobacco, Plant Physiol. 68, 808–813 (1981).
T. C. Taylor and I. Andersson, Structural transitions during activation and ligand binding in hexadecameric Rubisco inferred from the crystal structure of the activated unliganded spinach enzyme, Nature Struct. Biol. 3, 95–101 (1996).
A. P. Duff, T. J. Andrews, and P. M. G. Curmi, The transition between the open and closed states of Rubisco is triggered by the inter-phosphate distance of the bound bisphosphate, J. Mol. Biol. 298, 903–916 (2000).
T. C. Taylor and I. Andersson, The structure of the complex between Rubisco and its natural substrate ribulose 1,5-bisphosphate, J. Mol. Biol. 265, 432–444 (1997).
H. A. Schreuder, S. Knight, P. M. G. Curmi, et al., Formation of the active site of ribulose-1,5-bisphosphate carboxylase-oxygenase by a disorder-order transition from the unactivated to the activated form, Proc. Natl. Acad. Sci. USA 90, 9968–9972 (1993).
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Gao, F., Hong, F., Liu, C. et al. Mechanism of nano-anatase TiO2 on promoting photosynthetic carbon reaction of spinach. Biol Trace Elem Res 111, 239–253 (2006). https://doi.org/10.1385/BTER:111:1:239
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DOI: https://doi.org/10.1385/BTER:111:1:239