Regulation of ribulose 1,5-bisphosphate carboxylase-oxygenase activities by temperature pretreatment and chloroplast metabolites

doi:10.1016/0003-9861(76)90173-9

Archives of Biochemistry and Biophysics

Volume 176, Issue 1, September 1976, Pages 344-351

https://doi.org/10.1016/0003-9861(76)90173-9 Get rights and content

Abstract

Crystalline ribulose 1,5-bisphosphate carboxylase-oxygenase (EC 4.1.1.39) was isolated from tobacco (Nicotiana tabacum L.) leaf homogenates and the two competing reactions were examined for differential regulation in vitro by temperature pretreatment and chloroplast metabolites. Both the carboxylase and oxygenase activities were inactivated 50% by storing the dissolved protein at 0 °C and fully reactivated by heating the solution at 50 °C in the absence of Mg²⁺ and a sulfhydryl reagent. When the heat-activated enzyme was preincubated with physiological levels of various chloroplast metabolites and CO₂ and the two reactions were assayed simultaneously in the same reaction vessel upon initiation with ribulose 1,5-bisphosphate, three classes of effectors were observed: (a) those which stimulated both activities (NADPH, 6-phosphogluco-bisphosphate gluconate, fructose 1,6-bisphosphate, 3-phosphoglycerate glycerate), (b) those which essentially had no effect (fructose 6-phosphate, glucose 6-phosphate), and (c) one, ribose 5-phosphate, which inhibited the two reactions. However, within the limits of experimental error, none of the metabolites examined produced a differential regulation of the ribulose 1,5-bisphosphate carboxylase-oxygenase activities. The similar response of the two competing activities to temperature pretreatment and various chloroplast metabolites is consistent with the notion that both reactions are associated with the same or adjacent catalytic sites on this bifunctional enzyme.

References (37)

A. Weissbach et al.
J. Biol. Chem
(1956)
G. Bowes et al.
Biochem. Biophys. Res. Commun
(1971)
R. Chollet et al.
Biochem. Biophys. Res. Commun
(1975)
B.A. McFadden
Biochem. Biophys. Res. Commun
(1974)
G. Bowes et al.
J. Biol. Chem
(1972)
M.R. Badger et al.
Biochem. Biophys. Res. Commun
(1974)
N.E. Tolbert
B.B. Buchanan et al.
B.B. Buchanan et al.
J. Biol. Chem
(1973)
M.W. Kerr et al.
Phytohemistry
(1975)

G.D. Vogels et al.

Anal. Biochem

(1970)

N. Kawashima et al.

Biochem. Biophys. Res. Commun

(1971)

F.J. Ryan et al.

J. Biol. Chem

(1975)

T. Takabe et al.

Arch. Biochem. Biophys

(1975)

K. Lendzian et al.

Biochim. Biophys. Acta

(1975)

K. Werdan et al.

Biochim. Biophys. Acta

(1975)

F.J. Ryan et al.

J. Biol. Chem

(1975)

T.J. Andrews et al.

Biochemistry

(1973)

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The key enzyme of plant photosynthesis, d-ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco), must be activated to become catalytically competent via the carbamylation of Lys201 of the large subunit and subsequent stabilization by Mg²⁺ coordination. Many biochemical studies have reported that reduced nicotinamide adenine dinucleotide phosphate (NADPH) and 6-phosphogluconate (6PG) function as positive effectors to promote activation. However, the structural mechanism remains unknown. Here, we have determined the crystal structures of activated rice Rubisco in complex with NADPH, 6PG, or 2-carboxy-d-arabinitol 1,5-bisphosphate (2CABP). The structures of the NADPH and 6PG complexes adopt open-state conformations, in which loop 6 at the catalytic site and some other loops are disordered. The structure of the 2CABP complex is in a closed state, similar to the previous 2CABP-bound activated structures from other sources. The catalytic sites of the NADPH and 6PG complexes are fully activated, despite the fact that bicarbonate (NaHCO₃) was not added into the crystallization solution. In the catalytic site, NADPH does not interact with Mg²⁺ directly but interacts with Mg²⁺-coordinated water molecules, while 6PG interacts with Mg²⁺ directly. These observations suggest that the two effectors promote Rubisco activation by stabilizing the complex of Mg²⁺ and the carbamylated Lys201 with unique interactions and preventing its dissociation. The structure also reveals that the relaxed complex of the effectors (NADPH or 6PG), distinct from the tight-binding mode of 2CABP, would allow rapid exchange of the effectors in the catalytic sites by substrate d-ribulose 1,5-bisphosphate for catalysis in physiological conditions.
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After the transition of photosynthetic photon flux density (PPFD) from 700 μmol quanta m⁻² sec⁻¹ for one hour (step I) to 1400 μmol quanta m⁻² sec⁻¹ for one hour (step II) at air level of CO₂ and O₂, photosynthetic CO₂ fixation was suppressed to 60% in leaves of bean (Phaseolus vulgaris). Under these conditions, a similar extent of decrease occurred in both initial and total activities, and RuBP-binding free site concentration of ribulose 1, 5-bisphosphate car☐ylase-oxygenase whose protein content remained constant. Catalytic analysesin vitro indicated no significant deterioration of activation state of ribulose 1,5-bisphosphate car☐ylases with respect to Mg²⁺ and pH. Chloroplast metabolites including RUBP and 3-d-phosphoglycerate, whose stromal concentrations were slightly affected at step II, had a similar stimulatory and/or inhibitory effects on both activation and catalytic rates of ribulose 1,5-bisphosphate car☐ylases at steps I and II. Theoretical estimation does not totally account for the significant inhibition of RUBP car☐ylation reaction. The inhibition of photosynthetic rate, accompanied by a concomitant decrease in the free site concentration is considered to be due to a modified carbamylation state of the lysine residue at the active site, and a deteriorated activity of car☐yarabinitol 1-phosphate metabolism and RuBisCO activase, tightly related to a photosynthetic electron transport activity under a high PPFD for 1 hr inPhaseolus vulgaris leaves.
Ribulose 1,5-bisphosphate carboxylase-oxygenase I. Structural, immunochemical and catalytic properties
1987, Biochimie
Some structural, immunochemical and catalytic properties are examined for ribulose 1,5-bisphosphate carboxylase-oxygenase from various cellular organisms including bacteria, cyanobacteria, algae and higher plants. The native enzyme molecular masses and the subunit polypeptide compositions vary according to enzyme sources. The molecular masses of the large and small subunits from different cellular organisms, on the other hand, show a relatively high homology due to their well-conserved primary amino acid sequence, especially that of the large subunit. In higher plants, the native enzyme and the large subunit are recognized by the antibodies raised against either the native or large subunit, whereas the small subunit apparently cross-reacts only with the antibodies directed against itself. A wide diversity exists, however, in the serological response of the native enzyme and its subunits with antibodies directed against the native enzyme or its subunits from different cellular organisms. According to numerous kinetic studies, the carboxylase and oxygenase reactions of the enzyme with ribulose 1,5-bisphosphate and carbon dioxide or oxygen require activation by carbon dioxide and magnesium prior to catalysis with ribulose 1,5-bisphosphate and carbon dioxide or oxygen. The activation and catalysis are also under the regulation of other metal ions and a number of chloroplastic metabolites. Recent double-labeling experiments using radioactive ribulose 1,5-bisphosphate and ¹⁴CO₂ have elucidated the carboxylase/oxygenase ratios of the enzymes from different organisms. Another approach, i.e., genetic experiments, has also been used to examine the modification of the carboxylase/oxygenase ratio.
Quelques propriétés structurales, immunochimiques et catalytiques de la ribulose 1,5-bisphosphate carboxylase-oxygénase ont été examinées chez divers organismes cellulaires: bactéries, cyanobactéries, algues et végétaux supérieurs. La masse moléculaire de l'enzyme native et sa constitution en sous-unités varient selon les organismes cellulaires. Par contre, les masses moléculaires et les structures primaires de la grande et de la petite sous-unités sont relativement homogènes. La structure primaire de la grande sous-unité est en particulier très bien conservée. Chez les végétaux supérieurs, l'enzyme native et la grande sous-unité reconnaissent des anticorps dirigés contre l'une et l'autre, alors que la petite sous-unité paraît réagir uniquement avec des anticorps dirigés contre elle seule. Néanmoins, il existe chez différents organismes une grande diversité de réponse de l'enzyme et des sous-unités avec des anticorps. préparés contre chacune d'entre elles. De nombreuses études cinétiques ont montré que les réactions de carboxylation et d'oxygénation nécessitent l'activation préable de l'enzyme par le dioxide de carbone et le magnésium. L'activation et les fonctions catalytiques de l'enzyme sont également sous le contrôle d'autres ions métalliques et de nombreux métabolites chloroplastiques. Des étude double marquage en utilisant le ribulose 1,5-bisphosphate tritié et ¹⁴CO, ont permis de déterminer le rapport des activités carboxylase/oxygénase de l'enzyme chez différents organismes. Le rapport carboxylase/oxygénase a également pu être modifié par voie génétique.
Ribulose-1, 5-bisphosphate carboxylase from comfrey: Isolation and characterization
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Ribulose-1,5-bisphosphate carboxylase/oxygenase has been purified to electrophoretic homogeneity from comfrey, Symphytum spp. Sodium dodecyl sulfate polyacrylamide and polyacrylamide gel electrophoresis studies on the purified product showed no extraneous proteins. Comparisons of the electrophoretic mobilities of the subunits to those of standard proteins indicated a large subunit MW of 50 000 and a small subunit of 12 700, which for an octameric structure of each subunit indicates a native MW of 502 000. Specific activities of the comfrey enzyme ranged from 1.2 to nearly 2 μmol ¹⁴CO₂ fixed/min.mg of protein over several preparations and were maintained for months when stored from the sucrose gradient at − 70°. The specific activities depended critically on the amounts of enzyme used in the assay even under saturating conditions of substrates and cofactors. The effective pH dependence for carboxylase catalysis peaked near 7.4, which apparently is the lowest elective optimum yet reported for this enzyme from any source. However, on a constant carbon dioxide basis the pH dependence profile was reversed with a maximum near pH 8.6 which was 0.4 units higher than the value for the spinach enzyme. The K_ms for carbon dioxide and ribulose-1,5-bisphosphate at pH 7.5 were 130 μM and 30 μM, respectively, which are comparable to the accepted values for the carboxylase from spinach at pH 7.2.

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^☆: Central Research and Development Department Contribution No. 2370.

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Regulation of ribulose 1,5-bisphosphate carboxylase-oxygenase activities by temperature pretreatment and chloroplast metabolites☆

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