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1.
Anwaruzzaman ; Sawada Shinichi; Usuda Hideaki; Yokota Akiho 《Plant & cell physiology》1995,36(3):425-433
The mechanism of the regulation of the activation of ribulose1,5-bisphosphate carboxylase/ oxygenase (RuBisCO) by inorganicphosphate (Pi) in the presence of limiting concentrations ofCO2 was explored. The activation state of RuBisCO increasedsigmoidally following a biphasic kinetics against the concentrationof Pi in the activation mixture with an intermediary plateauat 2 to 3 mM Pi when the enzyme was activated for 30 min. Theintermediary plateau could not be seen when the preincubationtime was 10 min and the activation was completed at 10 mM Pi.RuBisCO from Euglena also showed a quite similar activationkinetics. The activation was not due to the contaminating CO2included in the stock Pi solution or in the activation buffercontaining the enzyme. The experiments with 2-carboxyarabinitol1,5-bisphosphate showed that the Pi stimulated activation wasdue to the promotion of binding of the activator CO2 to theactivation sites. It was also found that Pi increased the affinityof RuBisCO for the activator CO2 5.4-fold accompanied by a decreaseof the half-saturating concentration of CO2 to 1.6 µMat 20 mM MgCl2. Physiological significance of the effects ofPi on the activation of RuBisCO is discussed.
2Present address: Laboratory of Plant Molecular Physiology,Research Institute of Innovative Technology for the Earth (RITE),9-2 Kizugawadai, Kizu-cho, Soraku-gun, Kyoto, Japan. 相似文献
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Onizuka T Endo S Akiyama H Kanai S Hirano M Yokota A Tanaka S Miyasaka H 《Plant & cell physiology》2004,45(10):1390-1395
The operon encoding ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in the cyanobacterium Synechococcus sp. PCC7002 contains three rbc genes, rbcL, rbcX and rbcS, in this order. Introduction of translational frameshift into the rbcX gene resulted in a significant decrease in the production of large (RbcL) and small (RbcS) subunits of the Rubisco protein in Synechococcus sp. PCC7002 and in Escherichia coli. To investigate the function of the rbcX gene product (RbcX), we constructed the expression plasmid for the rbcX gene and examined the effects of RbcX on the recombinant Rubisco production in Escherichia coli. The coexpression experiments revealed that RbcX had marked effects on the production of large and small subunits of Rubisco without any significant influence on the mRNA level of rbc genes and/or the post-translational assembly of the Rubisco protein. The present rbcX coexpression system provides a novel and useful method for investigating the Rubisco maturation pathway. 相似文献
4.
Escherichia coli K-12 has the ability to migrate on semisolid media by means of swarming motility. A systematic and comprehensive collection of gene-disrupted E. coli K-12 mutants (the Keio collection) was used to identify the genes involved in the swarming motility of this bacterium. Of the 3,985 nonessential gene mutants, 294 were found to exhibit a strongly repressed-swarming phenotype. Further, 216 of the 294 mutants displayed no significant defects in swimming motility; therefore, the 216 genes were considered to be specifically associated with the swarming phenotype. The swarming-associated genes were classified into various functional categories, indicating that swarming is a specialized form of motility that requires a wide variety of cellular activities. These genes include genes for tricarboxylic acid cycle and glucose metabolism, iron acquisition, chaperones and protein-folding catalysts, signal transduction, and biosynthesis of cell surface components, such as lipopolysaccharide, the enterobacterial common antigen, and type 1 fimbriae. Lipopolysaccharide and the enterobacterial common antigen may be important surface-acting components that contribute to the reduction of surface tension, thereby facilitating the swarm migration in the E. coli K-12 strain. 相似文献
5.
Masakazu Inoue Tatsuko Egami Kanae Yokogawa Hirotada Kotani Toshio Morioka 《Bioscience, biotechnology, and biochemistry》2013,77(7):1391-1400
Cariogenic streptococci produce tenacious water-insoluble polysaccharides from sucrose and these form the structural intercellular matrix of dental plaque. Two Streptomyces species were isolated from soils on agar medium containing the water-insoluble polyglucan as a sole carbon source. They were identified as Streptomyces werraensis (strain F1) and Streptomyces chartreusis (strain F2). These strains produced extracellular enzymes which strongly solubilized the polyglucans from various strains of cariogenic streptococci. Strain F2 produced polyglucanases under rather stationary cultural condition, while F1 required vigorous aeration. The polyglucanases may provide a useful measure for the prevention and control of dental plaque formation, 相似文献
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Ascorbate peroxidase (APX) isoforms localized in the stroma and thylakoid membrane of chloroplasts play a central role in scavenging reactive oxygen species generated by photosystems. These enzymes are inactivated within minutes by H2O2 when the reducing substrate, ascorbate, is depleted. We found that, when the enzyme is inactivated by H2O2, a heme at the catalytic site of a stromal APX isoform is irreversibly cross-linked to a tryptophan residue facing the distal cavity. Mutation of this tryptophan to phenylalanine abolished the cross-linking and increased the half-time for inactivation from <10 to 62 s. In contrast with H2O2-tolerant peroxidases, rapid formation of the cross-link in APXs suggests that a radical in the reaction intermediate tends to be located in the distal tryptophan so that heme is easily cross-linked to it. This is the first report of a mutation that improves the tolerance of chloroplast APXs to H2O2. 相似文献
8.
Venkatesh B Arifuzzaman M Mori H Taguchi T Ohmiya Y 《Bioscience, biotechnology, and biochemistry》2004,68(10):2096-2103
The folding of many proteins including luciferase in vivo requires the assistance of molecular chaperone proteins. To understand how a chaperone targets luciferase, we took three luciferases that give different bioluminescence with the same luciferin substrate and with differences in homology. The three luciferase genes, firefly luciferase (FF-Luc) (from Pyrocoelia miyako), and red (RE-Luc) and green (GR-Luc) bioluminescence-emitting luciferases (from Phrixothrix railroad-worms), were expressed in Escherichia coli to produce fusion proteins with predicted molecular masses. Subsequently, we observed that DnaK and GroEL were co-purified along with recombinant luciferase. Although the amount of co-purified DnaK was almost the same compared to FF-Luc, GroEL was 25 and 32 times higher in GR-Luc and RE-Luc respectively. Furthermore, co-expression of GroEL/GroES along with luciferase substantially refolded RE-Luc and GR-Luc compared to FF-Luc. 相似文献
9.
Shimaoka T Ohnishi M Sazuka T Mitsuhashi N Hara-Nishimura I Shimazaki K Maeshima M Yokota A Tomizawa K Mimura T 《Plant & cell physiology》2004,45(6):672-683
A large number of proteins in the tonoplast, including pumps, carriers, ion channels and receptors support the various functions of the plant vacuole. To date, few proteins involved in these activities have been identified at the molecular level. In this study, proteomic analysis was used to identify new tonoplast proteins. A primary requirement of any organelle analysis by proteomics is that the purity of the isolated organelle needs to be high. Using suspension-cultured Arabidopsis cells (Arabidopsis Col-0 cell suspension), a method was developed for the isolation of intact highly purified vacuoles. No plasma membrane proteins were detected in Western blots of the isolated vacuole fraction, and only a few proteins from the Golgi and endoplasmic reticulum. The proteomic analysis of the purified tonoplast involved fractionation of the proteins by SDS-PAGE and analysis by LC-MS/MS. Using this approach, it was possible to identify 163 proteins. These included well-characterized tonoplast proteins such as V-type H+ -ATPases and V-type H+ -PPases, and others with functions reasonably expected to be related to the tonoplast. There were also a number of proteins for which a function has not yet been deduced. 相似文献
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