[30] Crystallization and assay procedures of tobacco ribulose-1,5-bisphosphate car☐ylase-oxygenase

Effects of Light and Elevated Atmospheric CO2 on the Ribulose Bisphosphate Carboxylase Activity and Ribulose Bisphosphate Level of Soybean Leaves

“…Furthermore, we report on substantial, light-induced variations in extractable RuBPCase activity from soybean leaves, which appear not to be mediated direct, natural solar irradiance (11 8.3 by the addition of 0.75 ml 2 M Tris base and 0.44 ml 4 N KOH. RuBP was then assayed using crystalline RuBPCase prepared from field-grown leaves of tobacco (Nicotiana tabacum cv NC25 12) as described by Kung et al (13). The assays were carried out at 30°C in a total volume of 1 ml in serum-stoppered Reacti-Vials which contained 0.4 ml of 50 mM Tris-HCl (pH 8.5), 10 mM MgCl2, 5 mM DTT, 20 mM NaH'4C03 (0.2 MCi/hmol), and a 0.5-ml aliquot of the supernatant leaf extract.…”

“…The assays were carried out at 30°C in a total volume of 1 ml in serum-stoppered Reacti-Vials which contained 0.4 ml of 50 mM Tris-HCl (pH 8.5), 10 mM MgCl2, 5 mM DTT, 20 mM NaH'4C03 (0.2 MCi/hmol), and a 0.5-ml aliquot of the supernatant leaf extract. The reaction was initiated with 0.1 ml of the tobacco RuBPCase (equivalent to 0.15 mg protein) which had been previously dissolved in 100 mm Tris-HCl, pH 8.5, 10 mm MgCl2, 10 mM NaHCO3, and 100 mM NaCl, and reactivated at 50°C for 25 min (13). After 1 h, the reaction was terminated with 0.1 ml 1 N HCI.…”

Relationship between the Kinetic Properties and the Small Subunit Composition of Nicotiana Ribulose-1, 5-bisphosphate Carboxylase

“…The kinetic properties of the genetically constructed hybrid RuBPCase molecules were determined from the crystallized enzyme using the standard procedure as described previously (10). A summary of the kinetic properties is presented in Table I.…”

Morphology and photosynthetic enzyme activity of maize phosphoenolpyruvate carboxylase transgenic rice

“…The carboxylase activity was assayed as described previously (Guo et al, 1988). For nicotinamide adenine dinucleotide phosphate (NADP)-malate dehydrogenase and ribulosebisphosphate carboxylase, the activity assay was separately performed using the methods described Li et al (1987) and Kung et al (1980).…”