The relative activities of PEP carboxylase and RuDP carboxylase in blue-green algae

The activity of phosphoenolpyruvate (PEP) carboxylase in preparations obtained by lysis of spheroplasts of three species of blue-green algae was found to be 1.5- to 5-fold higher at 20°C than that of RuDP carboxylase. This level of enzyme activity suggests that these algae may have the ability to fix carbon dioxide by a C₄ pathway.     

Inhibition of vitamin K-dependent carboxylase by metal ions and metal complexes: a reassessment

The vitamin K-dependent enzymatic carboxylation of glutamyl residues in blood protein precursors and in synthetic peptides is inhibited in vitro by transition metal complexes. Some authors suggested it is a result of metal ions interaction with intermediary oxygenated species. Using an oxygraph we have observed increases in the rate of oxygen utilization in the carboxylating system containing reduced vitamin K after addition of some transition metal ions and complexes. Kinetic studies indicate that, although oxygen utilization is increased by the addition of Cu2+, Fe3+, and hematin, the initial rate of carboxylation is not affected. The rate of carboxylation rapidly decreases at oxygen concentrations below 50 microM and reaches zero when oxygen is depleted. UV spectroscopy revealed simultaneous acceleration of the conversion of vitamin K hydroquinone into the parent quinone. The magnitude of these effects, as well as carboxylation inhibition, depends on the oxidation potential of the complexed ion and its lipophilicity. Addition of stable Mn parallel ion, which has no inhibitory effect on carboxylation, does not increase the rate of oxygen utilization nor the hydroquinone oxidation. The results suggest that inhibition of carboxylation by transition metals is mainly due to depletion of the necessary components (oxygen, vitamin K hydroquinone) of the carboxylating system rather than quenching of activated, oxygen-containing intermediates.     

Effects of manganese ions and magnesium ions on the activity of soya-bean ribulose bisphosphate carboxylase/oxygenase.

The Michaelis constants of soya-bean ribulose bisphosphate carboxylase for CO2 in the carboxylation reaction and for O2 in the oxygenation reaction depend on the nature of the bivalent cation present. In the presence of Mg2+ the Km for bicarbonate is 2.48 mM, and the Km for O2 is 37% (gas-phase concentration). With Mn2+ the values decrease to 0.85 mM and 1.7% respectively. For the carboxylation reaction Vmax. was 1.7 mumol/min per mg of protein with Mg2+ but only 0.29 mumol/min per mg of protein with Mn2+. For the oxygenation reaction, Vmax. values were 0.61 and 0.29 mumol/min per mg of protein respectively with Mg2+ and Mn2+.     

Effects of metal ions on activity of plasmin

The effects of some metal ions on amidolytic and fibrinogenolytic activities of highly purified human plasmin were investigated in vitro. In the presence of Zn²⁺, Cu²⁺, Cd²⁺, and Au⁺ in the incubation mixture at the concentrations of 1×10⁻⁵−1×10⁻³ M, the anidolytic plasmin activity was strongly inhibited, whereas Ca²⁺ and Mg²⁺ at the same concentrations were not effective. The analysis of the kinetic study has shown that Zn²⁺ or Cu²⁺ acts as mixed-type inhibitors of plasmin activity. The inhibition of amidolytic plasmin activity by Zn²⁺ and Cu²⁺ was reduced in the presence of EDTA, histidine, or albumin. Incubation of plasmin with Zn²⁺ or Cu²⁺ (at the concentration of 5×10⁻⁴ M) resulted in complete loss of its proteolytic action on fibrinogen, whereas Cd²⁺ and Au⁺ under the same conditions only partially inhibited this process.     

Effects of temperature on the cytokinin requirement of tobacco calluses

A cytokinin-nonrequiring strain (T22) was isolated from a cytokinin-requiringcallus strain T2 of tobacco (Nicotiana tabacum var. Bright Yellow). Strain T22 grew rapidly on the medium without added kinetinat 26?C. But its growth was completely suppressed at 16?C. Thisgrowth suppression at 16?C was partially recoverable by supplyingkinetin. Benzyladenine, geranylaminopurine and 2-methyl-8-benzylamino-s-triazolo[l,5-a]pyrazinewere also effective in removing growth suppression at 16?C.Adenine, which was unable to remove growth suppression of T22at 16?C, promoted the growth of T2 at 26?C, but not at 16?C.Physiological differences between cytokinin-requiring and -nonrequiringcalluses are discussed. ¹Part II in the series "Studies, on Plant Tissue Cultures";for Part I, See Plant & Cell Physiol. 9: 103–114 (1968). (Received May 29, 1970; )     

Changes in phosphoenolpyruvate carboxylase and ribulosebisphosphate carboxylase activities in tobacco plants infected with tobacco mosaic virus

Considerable changes in the activities of phosphoenolpyruvate carboxylase and ribulosebisphosphate carboxylase were found inNicotiana tabacum cv. Sarasun plants infected with TMV. Ribulosebisphosphate carboxylase is inhibited at the time of maximum TMV reproduction, but its decreased activity is at the same time partly compensated by phosphoenol-pyruvate carboxylase in the shoots of infected plants. The pattern of activity of this enzyme nearly exactly reflects the pattern of reproduction of the tobacco mosaic virus.     

Ordered association of tobacco mosaic virus in the presence of divalent metal ions

The formation of ordered aggregates of tobacco mosaic virus (TMV) in the presence of divalent metal ions has been studied in concentrated (1-25 mg/ml) solutions of the virus. The divalent metal cations Cd2+, Zn2+, Pb2+, Cu2+, and Ni2+ have been found to promote TMV precipitation from solution at a critical concentration Ccrit, which for a given metal depends on the pH and the ionic strength of the solution, but is largely independent of the virus concentration. The TMV precipitate behaves as a nematic liquid crystal and on drying at a glass surface produces highly ordered, optically birefringent films. However, precipitation is not observed with alkali-earth metals such as Ca2+ and Mg2+. The experimental data suggest that, apart from two 'internal' metal-binding sites in each TMV subunit, the virus contains metal-binding sites of a lower affinity which promote cross-linking of TMV rods via metal bridges. The latter seem to be responsible for the precipitation of TMV in the presence of divalent cations at neutral pH. We propose that the metal-induced cross-linking may be the predominant mechanism to account for the limited solubility of a variety of proteins in solution containing metal cations with valence 2 and higher.     

Effects of magnesium, manganese and adenosine triphosphate ions on pyruvate carboxylase from baker''s yeast

1. Pyruvate carboxylase from baker's yeast acts with either MgATP(2-) or MnATP(2-) as substrate. The optimum pH for the enzyme reaction is 8.0 with MgATP(2-) and 7.0 with MnATP(2-). 2. When the reaction velocity is plotted against MgATP(2-) (or MnATP(2-)) concentration slightly sigmoid curves are obtained, either in the presence or in the absence of acetyl-CoA (an allosteric activator). In the presence of excess of free Mg(2+) (or Mn(2+)) the curves turn into hyperbolae, whereas in the presence of excess of free ATP(4-) the apparent sigmoidicity of the curves increases. 3. The sigmoidicity of the plots of v against MgATP(2-) (or MnATP(2-)) concentration can be explained by the inhibitory effect of free ATP(4-), the concentration of which, in the experimental conditions employed, is significant and varies according to the total concentration of the ATP-magnesium chloride (or ATP-manganese chloride) mixture. Free ATP(4-) behaves as a negative modifier of yeast pyruvate carboxylase. 4. The effect of high concentrations of Mg(2+) (or Mn(2+)) on the kinetics of yeast pyruvate carboxylase can be explained as a deinhibition with respect to ATP(4-), instead of a direct enzyme activation. 5. At pH6.5 manganese chloride is more effective than magnesium chloride as enzyme activator even in the presence of a great excess (16-fold) of the latter. This is consistent with a significant contribution of the MnATP(2-) complex to the activity of yeast pyruvate carboxylase, in medium conditions resembling those existing inside the yeast cell (pH6.25-6.75; 12mm-magnesium chloride and 0.75mm-manganese chloride). 6. The physiological significance of the enzyme inhibition by free ATP(4-) is doubtful since the Mg(2+) and Mn(2+) concentrations reported to exist inside the yeast cell are sufficient to decrease ATP(4-) concentrations to ineffective values.     

Effects of pH and metal ions on antioxidative activities of catechins

The Effects of pH on antioxidative activities of catechol, pyrogallol, and four catechins, and effects of metal ions (Al3+, Ca2+, Cd2+, Co2+, Cr3+, Cu2+, Fe2+, Fe3+, K+, Mg2+, Mn2+, Na+, and Zn2+) on antioxidative activities of (-)-epigallocatechin gallate (EGCG) were studied by an oxygen electrode method. The antioxidative activities of catechins were high and constant at pH 6-12, but decreased in acidic and strong alkaline solutions. Copper(II) ion the most strongly increased the antioxidative activity of EGCG among these metal ions examined, but iron(II) ion largely inhibited the antioxidative activity of EGCG. These effects are discussed considering the formation of metal complexes with catechins and the change in oxidation potentials.     

Effects of metal ions on the structure and spectra of the peptide group.

The interaction between salts of Groups IA and IIA and model aliphatic amides has been studied. The interaction has been monitored by calorimetry and spectroscopy. Among the alkali cations, lithium ion interacts strongest, while in Group II calcium appears to interact somewhat stronger than magnesium. The metal cation binds to the amide oxygen atom and causes alteration in the amide group geometry. As a consequence, significant alterations are seen in the infrared, nuclear magnetic resonance, ultraviolet, and circular dichroism spectra of the ligand peptide. These findings are suggested to be of importance to the conformational studies of polypeptides and proteins.     

The titration curve of insulin in the presence of various bivalent metal ions

1. Titration curves of insulin in the presence and absence of various metal ions are reported. 2. The difference in base consumption with and without the metal ions is compared with calculated curves. 3. These experiments suggest that in dilute solutions Zn(2+) and Cu(2+) ions are bound to alpha-amino groups.     

Effects of metal ions on sphingomyelinase activity of Bacillus cereus

Some divalent metal ions were examined for their effects on sphingomyelinase activity of Bacillus cereus. The enzyme activity toward mixed micelles of sphingomyelin and Triton X-100 proved to be stimulated by Co2+ and Mn2+, as well as by Mg2+. Km's for Co2+ and Mn2+ were 7.4 and 1.7 microM, respectively, being smaller than the Km for Mg2+ (38 microM). Sr2+ proved to be a competitive inhibitor against Mg2+, with a Ki value of 1 mM. Zn2+ completely abolished the enzyme activity at concentrations above 0.5 mM. The concentration of Zn2+ causing 50% inhibition of the enzyme activity was 2.5 microM. Inhibition by Zn2+ was not restored by increasing concentrations of Mg2+ when the concentration of Zn2+ was above 10 microM. Ba2+ was without effect. When sphingomyelinase was incubated with unsealed ghosts of bovine erythrocytes at 37 degrees C, the enzyme was significantly adsorbed onto the membrane in the presence of Mn2+, Co2+, Sr2+ or Ba2+. Incubation with intact or Pronase-treated erythrocytes caused enzyme adsorption only in the presence of Mn2+. In the course of incubation, the enzyme was first adsorbed on the membranes of intact bovine erythrocytes in the presence of Mn2+; then sphingomyelin breakdown proceeded with ensuing desorption of adsorbed enzyme. Hot-cold hemolysis occurred in parallel with sphingomyelin breakdown. In this case, the hydrolysis of membranous sphingomyelin as well as the initial enzyme adsorption took place in the following order: unsealed ghosts greater than Pronase-treated erythrocytes greater than intact erythrocytes.     

金属离子对生物絮凝剂REA-11合成的影响

考察了Mg2 、Na 、K 、Ca2 对谷氨酸棒杆菌生长及生物絮凝剂合成的影响。结果表明,培养基中添加适当的Mg2 对菌体的生长和絮凝剂的产生有一定的促进作用。K 和Ca2 对絮凝剂的产生具有一定的抑制作用。Na 的添加对菌体的生长和絮凝剂的产生均无显著影响。     

Antibacterial activity of chitosan tripolyphosphate nanoparticles loaded with various metal ions

The aim of this study was to prepare and select chitosan nanoparticles loaded metal ions with high antibacterial activities. Chitosan nanoparticles were prepared based on ionic gelation between chitosan and sodium tripolyphosphate. Then, Ag⁺, Cu²⁺, Zn²⁺, Mn²⁺, or Fe²⁺ was individually loaded onto chitosan nanoparticles. Their particle sizes and zeta potentials were measured. Their antibacterial activities were evaluated by determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Escherichia coli 25922, Salmonella choleraesuis ATCC 50020 and Staphylococcus aureus 25923 in vitro. Results showed that antibacterial activity was significantly enhanced by the metal ions loaded, except for Fe²⁺. Especially for chitosan nanoparticles loaded Cu²⁺, the MIC and MBC against E. coli 25922, S.choleraesuis ATCC 50020 and S. aureus 25923 were 21–42 times lower than that of Cu²⁺, respectively. Moreover, it was found that antibacterial activity was directly proportional to zeta potential.