Effect of human neuronal tau on denaturation and reactivation of rabbit muscle D-glyceraldehyde-3-phosphate dehydrogenase

2000 ◽  
Vol 351 (1) ◽  
pp. 233-240 ◽  
Author(s):  
Yong-Hui CHEN ◽  
Rong-Qiao HE ◽  
Ying LIU ◽  
Yang LIU ◽  
Zhi-Gang XUE

Human neuronal tau-40 (htau-40) has been used to study denaturation and renaturation of rabbit muscle D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12). Inactivation of GAPDH incubated with tau was more distinguishably detected than that of control GAPDH during thermal and guanidine hydrochloride (GdnHCl) denaturation. However, tau did not influence the activity of GAPDH at room temperature or in solution without GdnHCl. A marked change in both the emission intensity and emission maximum of the intrinsic fluorescence at 335nm of GAPDH with tau was observed when GdnHCl concentration was 0.8M, but that of the control without tau occurred in 1.2M GdnHCl. The first-order rate of the decrease in the fluorescence intensity of the enzyme with tau was approximately twice as great as that of GAPDH without tau. Kinetics of inactivation of GAPDH with tau in 0.2M GdnHCl was a monophasic procedure, instead of the biphasic procedure followed by the control, as described before [He, Zhao, Yan and Li (1993) Biochim. Biophys. Acta 1163, 315–320]. Similar results were obtained when the enzyme was thermally denatured at 45°C. It revealed that tau bound to the denatured GAPDH but not the native molecule. On the other hand, tau suppressed refolding and reactivation of GAPDH when this enzyme was reactivated by dilution of GdnHCl solution. Furthermore, tau improved the aggregation of the non-native GAPDH in solutions. It suggested that tau acted in an anti-chaperone-like manner towards GAPDH in vitro. However, tau lost that function when it was aggregated or phosphorylated by neuronal cdc2-like protein kinase. It showed that tau's anti-chaperone-like function depended on its native conformation.

1992 ◽  
Vol 57 (11) ◽  
pp. 2302-2308
Author(s):  
Karel Mocek ◽  
Erich Lippert ◽  
Emerich Erdös

The kinetics of the reaction of solid sodium carbonate with sulfur dioxide depends on the microstructure of the solid, which in turn is affected by the way and conditions of its preparation. The active form, analogous to that obtained by thermal decomposition of NaHCO3, emerges from the dehydration of Na2CO3 . 10 H2O in a vacuum or its weathering in air at room temperature. The two active forms are porous and have approximately the same specific surface area. Partial hydration of the active Na2CO3 in air at room temperature followed by thermal dehydration does not bring about a significant decrease in reactivity. On the other hand, if the preparation of anhydrous Na2CO3 involves, partly or completely, the liquid phase, the reactivity of the product is substantially lower.


1996 ◽  
Vol 313 (1) ◽  
pp. 45-50 ◽  
Author(s):  
Alexander V. SKURAT ◽  
Peter J. ROACH

Glycogen synthase can be inactivated by sequential phosphorylation at the C-terminal residues Ser652 (site 4), Ser648 (site 3c), Ser644 (site 3b) and Ser640 (site 3a) catalysed by glycogen synthase kinase-3. In vitro, glycogen synthase kinase-3 action requires that glycogen synthase has first been phosphorylated at Ser656 (site 5) by casein kinase II. Recently we demonstrated that inactivation is linked only to phosphorylation at site 3a and site 3b, and that, in COS cells, modification of these sites can occur by alternative mechanisms independent of any C-terminal phosphorylations [Skurat and Roach (1995) J. Biol. Chem. 270, 12491-12497]. To address these mechanisms multiple Ser → Ala mutations were introduced in glycogen synthase such that only site 3a or site 3b remained intact. Additional mutation of Arg637 → Gln eliminated phosphorylation of site 3a, indicating that Arg637 may be important for recognition of site 3a by its corresponding protein kinase(s). Similarly, additional mutation of Pro645 → Ala eliminated phosphorylation of site 3b, indicating a possible involvement of ‘proline-directed’ protein kinase(s). Mutation of Arg637 alone did not activate glycogen synthase as expected from the loss of phosphorylation at site 3a. Rather, mutation of both Arg637 and the Ser → Ala substitution at site 3b was required for substantial activation. The results suggest that sites 3a and 3b can be phosphorylated independently of one another by distinct protein kinases. However, phosphorylation of site 3b can potentiate phosphorylation of site 3a, by an enzyme such as glycogen synthase kinase-3.


1977 ◽  
Vol 55 (19) ◽  
pp. 2530-2534 ◽  
Author(s):  
F. Maillard ◽  
J.-P. Zrÿd

Incubation of cell suspensions of sycamore (Acer pseudoplatanus) with β-indoyl-3-acetic acid (IAA) first led to the formation of IAA-glycosides, then to that of IAA-aspartate. Great differences are observed between the kinetics of IAA transformed by two distinct strains: one, auxin dependent (S), the other, auxin independent (MB). Other degradation products are only found in the culture medium. The localization of IAA-degrading systems in the cell wall is postulated. The auxin requirement of the S strain is discussed.


1938 ◽  
Vol 16b (5) ◽  
pp. 176-193 ◽  
Author(s):  
E. W. R. Steacie ◽  
I. E. Puddington

The kinetics of the thermal decomposition of n-butane has been investigated at pressures from 5 to 60 cm. and temperatures from 513 to 572 °C. The initial first order rate constants at high pressures are given by[Formula: see text]The results are in good agreement with the work of Frey and Hepp, but differ greatly from that of Paul and Marek. The reaction rate falls off strongly with diminishing pressure; this is rather surprising for a molecule as complex as butane. The first order constants in a given run fall rapidly as the reaction progresses. The last two facts suggest that chain processes may be involved.A large number of analyses of the products of reaction have been made at various pressures, temperatures, and stages of the reaction, the method being that of low-temperature fractional distillation. The products are virtually independent of temperature and pressure over the range investigated. The initial products, obtained by extrapolation to zero decomposition, are:—H2, 2.9; CH4, 33.9; C3H6, 33.9; C2H4, 15.2; C2H6, 14.1%. The mechanism of the reaction is discussed, and the results are compared with those of the other paraffin decompositions.


2011 ◽  
Vol 399-401 ◽  
pp. 1067-1070
Author(s):  
Chun Yan Li ◽  
Cong Cong Hu ◽  
Zhi Guo Wen ◽  
Sheng Xiong Dong

The method of high performance liquid chromatography (HPLC) is established to determine the content of antibacterial agent — ciprofloxacin (CF) in the degradation solution of ciprofloxacin-polyurethane (CFPU) and investigate the in vitro degradation kinetics by plotting and fitting the cumulative release curves to inspect the effects of different medium and different concentrations on drug release. The results showed that the HPLC method is accurate, reliable and simple. The drug-release of CFPU was bioresponsive and could be accorded with first order kinetics. It was observed that CF was released from CFPU by a combination of diffusion and erosion mechanism, mainly in the manner of diffusion in the absence of infection while erosion mechanism in the presence of infection.


1958 ◽  
Vol 36 (12) ◽  
pp. 1729-1734 ◽  
Author(s):  
J. E. Hazell ◽  
K. E. Russell

The reaction of DPPH (2,2-diphenyl-1-picrylhydrazyl) with N-phenyl-1-naphthylamine, N-phenyl-2-naphthylamine, diphenylamine, and methylaniline has been studied and has been shown to be primarily a hydrogen abstraction process. Two moles DPPH react with 1–1.15 moles secondary amine to give 1.7–1.8 moles 2,2-diphenyl-1-picrylhydrazine and further products.The reaction between DPPH and N-phenyl-1-naphthylamine is first order with respect to each reactant. The reaction of DPPH with the other amines is retarded by the major product 2,2-diphenyl-1-picrylhydrazine and the kinetics of the over-all reaction are complex. However second-order rate constants and activation energies have been obtained using initial rates of reaction. Possible reaction mechanisms are discussed.


2021 ◽  
Vol 33 (8) ◽  
pp. 1929-1934
Author(s):  
P. Gupta ◽  
A.K. Verma ◽  
P. Chaudhuri (Chattopadhyay)

Maintenance of native structure and function of the protein is a major concern for industrial production of aggregation prone therapeutically important recombinant proteins. Aggregation may results due to change in the native conformation of proteins under different stress conditions. To overcome the problem of protein aggregation, role of silver and gold nanoparticles have been investigated. The nanoparticles owing to their affirmative interaction with the proteins possess chaperoning activities and protect the native state from denaturation. In the present study, through performing chemical denaturation of zebrafish dihydrofolate reductase using denaturants like guanidine hydrochloride and urea in the presence and absence of gold and silver nanoparticles and monitoring the process through enzyme activity assay and intrinsic tryptophan fluorescence, we have demonstrated the impact of nanoparticles in maintaining native conformation of proteins. Further, the outcome of refolding studies of DHFR protein with nanoparticles monitored by UV-visible spectroscopy was also reported.


2020 ◽  
Vol 9 (4) ◽  
pp. e107942900
Author(s):  
Alvaro Gustavo Ferreira da Silva ◽  
Franciscleudo Bezerra da Costa ◽  
Yasmin Lima Brasil ◽  
Brencarla de Medeiros Lima ◽  
Eder Pereira da Rocha Sousa ◽  
...  

Ziziphus joazeiro is endemic to the brazilian Caatinga and its fruits can be used as a food supplement by accelerating the ethanol metabolism in the body and reducing the alcohol harmful effects due to high alcohol dehydrogenase (ADH) activity. The objective was to determine the kinetics of ADH activity, in different incubation times, of Z. joazeiro mature fruits as a food supplement. Ziziphus joazeiro fruits, at the fourth maturation stage, were incubated for 0 (no incubation), 3, 6, 12, 24 and 48 hours at room temperature. The ADH activity was determined. ADH activity was higher in fruits incubated for 0 and 3 h. The ADH activity was higher in the early incubation times, probably due to the greater availability of NAD+, which after being reduced to NADH delayed regeneration. Without the cofactor in the oxidized form, the enzymatic activity decreases. Ziziphus joazeiro fruit has the potential to be used as a food supplement accelerating the alcohol metabolism and reducing the harmful effects.


1987 ◽  
Vol 52 (6) ◽  
pp. 1527-1544 ◽  
Author(s):  
Ulrika Králíková ◽  
Martin Bajus ◽  
Jozef Baxa

The kinetics of pyrolysis of methylcyclohexane was investigated from the viewpoint of coke formation in a steel tubular reactor (S/V = 6·65 cm-1) at 0·1 MPa, 700 to 820 °C and residence time 0·01 to 0·24 s. Decomposition of methylcyclohexane proceeds as a first order reaction with a frequency factor 6·31 . 1015 s-1 and activation energy 251·2 kJ mol-1. The course of secondary reactions associated with the formation of coke is discussed. Investigation of coke formation showed a greater tendency of methylcyclohexane to coking in comparison with heptane. A prominent role plays the course of dehydrogenation of cycloalkane radicals up to aromates, this being reflected by the overall conversion of methylcyclohexane, and, on the other hand the thus formed aromates enter the consecutive reactions leading to coke.


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