scholarly journals Reactivation of triosephosphate isomerase from three trypanosomatids and human: effect of Suramin

1998 ◽  
Vol 332 (1) ◽  
pp. 91-96 ◽  
Author(s):  
Xiu-Gong GAO ◽  
Georgina GARZA-RAMOS ◽  
Emma SAAVEDRA-LIRA ◽  
Nallely CABRERA ◽  
Marietta T. de GÓMEZ-PUYOU ◽  
...  
Keyword(s):  
Protein Concentration ◽  
Triosephosphate Isomerase ◽  
Guanidine Hydrochloride ◽  
Monomer Concentration ◽  
Life Time ◽  
The Other ◽  
Drug Of Choice ◽  
Rate Limiting Step ◽  
Order Of Magnitude ◽  
Rate Limiting

The reactivation of the homodimeric triosephosphate isomerases (TIMs) from Trypanosoma brucei, T. cruzi, Leishmania mexicana and humans was determined after their denaturation with guanidine hydrochloride. In the range of 2–32 µg of T. brucei TIM per ml and 0.2–5 µg of the other enzymes per ml, the rate and extent of TIM reactivation depended on protein concentration, indicating that at these protein concentrations, the rate-limiting step of reactivation is monomer association and not monomer folding. The rate of monomer association was more than one order of magnitude lower in the T. brucei enzyme than in the other three enzymes. Suramin is a drug of choice in the treatment of sleeping sickness, but its mechanism of action is not known. At micromolar concentrations, Suramin inhibited the reactivation of the four enzymes, but the extent of inhibition by Suramin decreased with increasing protein concentration as consequence of a diminution of the life time of the folded monomer. Since the life time of the monomer of T. brucei TIM is longer than that of the other enzymes, Suramin is a more effective inhibitor of the reactivation of TIM from T. brucei, particularly at monomer concentrations above 1 µg of protein per ml (monomer concentration approx. 37 nM). Compounds that are structurally related to Suramin also inhibit TIM reactivation; their effect was about five times more pronounced in the enzyme from T. brucei than in human TIM.

Distribution of ornithine decarboxylase in ovaries of rat and hamster during pro-oestrus

1986 ◽  
Vol 113 (3) ◽  
pp. 403-409 ◽  
Author(s):  
Lo Persson ◽  
Karin Isaksson ◽  
Elsa Rosengren ◽  
Frank Sundler
Keyword(s):  
Ornithine Decarboxylase ◽  
Granulosa Cells ◽  
Physiological Function ◽  
The Other ◽  
Interstitial Tissue ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Other Hand ◽  
Rat Ovary ◽  
Rate Limiting

Abstract. The biosynthesis of polyamines is dramatically increased in the ovaries of rat and hamster during the evening of pro-oestrus. In an attempt to shed some light on the physiological function of this biosynthesis ornithine decarboxylase (ODC), which catalyzes the rate-limiting step in the biosynthesis of the polyamines, was immunohistochemically localized in the ovaries from rat and hamster during pro-oestrus. At dioestrus, only a few immunoreactive cells were found in the ovaries. During the evening of pro-oestrus, on the other hand, numerous immunoreactive cells were observed in the ovaries. These cells were confined to the internal thecal layer of Graafian as well as smaller follicles and to the interstitial tissue of the ovary. The granulosa cells appeared to be devoid of immunoreactive ODC. The hamster ovary, which during this time exhibited considerably higher levels of ODC activity than the ovaries from the rat, did accordingly contain more immunoreactive cells than the rat ovary.

Reactions of 1,3-diacylthioureas with methoxide ion and with amines

1987 ◽  
Vol 52 (1) ◽  
pp. 120-131 ◽  
Author(s):  
Jaromír Kaválek ◽  
Josef Jirman ◽  
Vojeslav Štěrba
Keyword(s):  
Carbonyl Group ◽  
Rate Constants ◽  
Dissociation Constants ◽  
Equilibrium Constants ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Order Of Magnitude ◽  
Acetyl Group ◽  
Methoxide Ion ◽  
Rate Limiting

Rate constants of base-catalyzed methanolysis and dissociation constants in methanol have been determined for benzoylthiourea (II), 1,3-diacetylthiourea (III), 1,3-dibenzoylthiourea (IV), and 1-acetyl-3-benzoylthiourea (V). With the diacyl derivatives III and IV, the reaction of methoxide ion with the neutral substrate is accompanied by that of methoxide with the substrate anion (at higher alkoxide concentrations). Above 0.1 mol l-1 CH3O(-), the rate constants are also affected by medium. The rate of the reaction of neutral diacyl derivative is decreased, and that of the reaction of methoxide with the substrate anion is rapidly increased. The dissociation constant of II is higher than that of acetylthiourea (I) by about one order of magnitude, but the attack of methoxide on the carbonyl group of II is about three times slower than that in I. The benzoyl group at the N1 nitrogen exhibits a greater activating influence (in both the rate and the equilibrium constants) on the other NHCOR group than the acetyl group does. With V the ratio of methanolysis rate constants is 9 : 1 in favour of the acetyl group. The reaction of diacetyl derivative III with 1-butanamine has been followed in butanamine buffers. At the lowest butanamine concentrations, the reaction is second order in the amine, and the rate-limiting step is the proton transfer from the intermediate to the second amine molecule. At the highest butanamine concentrations the reaction becomes first order in the amine, and the rate-limiting step changes to the attack of butanamine on the carbonyl group of diacetyl derivative III.

Kinetics and mechanism of methanolysis of benzoyl derivatives of substituted phenylureas and phenylthioureas

1984 ◽  
Vol 49 (9) ◽  
pp. 2103-2110 ◽  
Author(s):  
Jaromír Kaválek ◽  
Said El Bahaie ◽  
Vojeslav Štěrba
Keyword(s):  
Nitrogen Atom ◽  
Steric Hindrance ◽  
Rate Constants ◽  
Dissociation Constants ◽  
Kinetics And Mechanism ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Order Of Magnitude ◽  
Rate Limiting ◽  
Derivatives Of

The methanolysis rate constants and dissociation constants have been measured of benzoyl derivatives of substituted phenylureas and phenylthioureas. The dissociation constants of the thio derivatives are higher by 1 order of magnitude and the rate constants are higher by 2 orders of magnitude than the respective values of the oxygen analogues. Logarithms of the rate and dissociation constants have been correlated with the Hammet σ constant; the ρ constant of the methanolysis of the oxygen derivatives is almost 2x higher than that of the thio derivatives, which is explained by a change in the rate-limiting step. Methylation of the phenyl nitrogen atom increases the acidity by almost 2 orders of magnitude. This effect is due obviously to steric hindrance to the conjugation with the adjacent carbonyl or thiocarbonyl group.

scholarly journals Block of current through single calcium channels by Fe, Co, and Ni. Location of the transition metal binding site in the pore.

1991 ◽  
Vol 97 (2) ◽  
pp. 351-367 ◽  
Author(s):  
B D Winegar ◽  
R Kelly ◽  
J B Lansman
Keyword(s):  
Metal Binding ◽  
The Other ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Coulombic Interactions ◽  
Rate Limiting Step ◽  
Ion Size ◽  
Rate Limiting ◽  
Kinetics Of ◽  
Ca2 Channels

The blocking actions of Fe2+, Co2+, and Ni2+ on unitary currents carried by Ba2+ through single dihydropyridine-sensitive Ca2+ channels were recorded from cell-attached patches on myotubes from the mouse C2 cell line. Adding millimolar concentrations of blocker to patch electrodes containing 110 mM BaCl2 produced discrete excursions to the closed channel level. The kinetics of blocking and unblocking were well described with a simple model of open channel block. Hyperpolarization speeded the exit of all of the blockers from the channel, as expected if the blocking site resides within the pore. The block by Ni2+ differs from that produced by Fe2+ and Co2+ because Ni2+ enters the channel approximately 20 times more slowly and exits approximately 50 times more slowly. Ni2+ also differs from the other transition metals because at millimolar concentrations it reduces the amplitude of the unitary current in a concentration-dependent manner. The results are consistent with the idea that the rate-limiting step for ion entry into the channel is water loss at its inner coordination sphere; unblocking, on the other hand, cannot be explained in terms of simple coulombic interactions arising from differences in ion size.

scholarly journals Importance of catalase in the disposal of hydrogen peroxide within human erythrocytes

Blood ◽  
1994 ◽  
Vol 84 (1) ◽  
pp. 325-330 ◽  
Author(s):  
GF Gaetani ◽  
HN Kirkman ◽  
R Mangerini ◽  
AM Ferraris
Keyword(s):  
Glutathione Peroxidase ◽  
Human Erythrocytes ◽  
The Other ◽  
Unexpected Result ◽  
System Function ◽  
Sequential Action ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Genetic Deficiency ◽  
Rate Limiting

The catalase within normal, intact human erythrocytes was completely inactivated with amino triazole. The rate of 14CO2 evolution, when the cells were subsequently incubated with 14C-labeled glucose, provided a measure of the rate at which NADPH was being oxidized by the glutathione peroxidase/reductase system for the disposal of H2O2. This rate was determined in control cells and in catalase-inactivated cells while the cells were exposed to H2O2, which was generated at various constant and predetermined rates by glucose oxidase. The results indicated that catalase handles approximately half of the generated H2O2. The glutathione peroxidase/reductase mechanism accounted for the other half. These results are in agreement with our earlier findings on erythrocytes of a subject with a genetic deficiency of catalase. However, an unexpected result with the present approach was the finding that the increased dependence on the glutathione peroxidase/reductase mechanism did not occur until greater than 98% of the catalase had been inactivated. The latter observation indicates that catalase and the glutathione peroxidase/reductase system function intracellularly in a manner very different from that previously ascribed to them. An explanation of the findings requires that the two methods of H2O2 disposal function in a coordinated way, such as a sequential action in which the glutathione peroxidase/reductase system is the rate-limiting step.

scholarly journals Modulation of inhibition of ferrochelatase by N-methylprotoporphyrin

2006 ◽  
Vol 399 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Zhen Shi ◽  
Gloria C. Ferreira
Keyword(s):  
Protoporphyrin Ix ◽  
Kinetic Mechanism ◽  
Wild Type ◽  
Rate Limiting Step ◽  
Transition State Analogue ◽  
Mammalian Cell Cultures ◽  
Kinetic Mechanisms ◽  
Specificity Constant ◽  
Order Of Magnitude ◽  
Rate Limiting

Protoporhyrin IX ferrochelatase catalyses the terminal step of the haem-biosynthetic pathway by inserting ferrous iron into protoporphyrin IX. NMPP (N-methylprotoporphyrin), a transition-state analogue and potent inhibitor of ferrochelatase, is commonly used to induce haem deficiency in mammalian cell cultures. To create ferrochelatase variants with different extents of tolerance towards NMPP and to understand further the mechanism of ferrochelatase inhibition by NMPP, we isolated variants with increased NMPP resistance, bearing mutations in an active-site loop (murine ferrochelatase residues 248–257), which was previously shown to mediate a protein conformational change triggered by porphyrin binding. The kinetic mechanisms of inhibition of two variants, in which Pro255 was replaced with either arginine (P255R) or glycine (P255G), were investigated and compared with that of wild-type ferrochelatase. While the binding affinity of the P255X variants for NMPP decreased by one order of magnitude in relation to that of wild-type enzyme, the inhibition constant increased by approximately two orders of magnitude (Kiapp values of 1 μM and 2.3 μM for P255R and P255G respectively, as against 3 nM for wild-type ferrochelatase). Nonetheless, the drastically reduced inhibition of the variants by NMPP was not paralleled with a decrease in specificity constant (kcat/Km, protoporhyrin IX) and/or catalytic activity (kcat). Further, although NMPP binding to either wild-type ferrochelatase or P255R occurred via a similar two-step kinetic mechanism, the forward and reverse rate constants associated with the second and rate-limiting step were comparable for the two enzymes. Collectively, these results suggest that Pro255 has a crucial role in maintaining an appropriate protein conformation and modulating the selectivity and/or regiospecificity of ferrochelatase.

Propargylic and Chromene Terminated Prepolymers. 3. Study of Homopolymerization and the Effect of the Chromene Content on Reaction Kinetics

1996 ◽  
Vol 8 (3) ◽  
pp. 341-361 ◽  
Author(s):  
M F Grenier-Loustalot ◽  
C Sanglar
Keyword(s):  
Reaction Kinetics ◽  
Rate Constant ◽  
Reaction Temperature ◽  
Considerable Effect ◽  
The Other ◽  
Thermally Induced ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Physicochemical Data ◽  
Rate Limiting

We have chemically synthesized CTR (chromene terminated resin) prepolymers in order to more specifically study the homopolymerization reaction of chromene. Physicochemical data were used to show the value of working with dichromene prepolymers instead of propargylic monomers. In particular, the homopolymerization reaction is less exothermic than the thermally induced ring formation. So, when the homopolymerization reaction is predominant, the processability of the final thermoset material will be easier. Besides, the rate constant of the homopolymerization reaction is higher than that measured for dipropargylic monomers. This explains the interest in overcoming β stage formation which is the kinetically rate limiting step. On the other end, the presence of residual propargylic functions in dichromene prepolymers after synthesis has no considerable effect on reaction temperature or advancement of reaction.

Effect of protein concentration on the formation of glycated albumin and fructosamine

1991 ◽  
Vol 37 (12) ◽  
pp. 2138-2139 ◽  
Author(s):  
E Lamb ◽  
R Mainwaring-Burton ◽  
A Dawnay
Keyword(s):  
Protein Concentration ◽  
Glycated Albumin ◽  
Open Chain ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Considerable Debate ◽  
Lysine Residues ◽  
Available Lysine ◽  
Rate Limiting ◽  
Serum Protein Concentration

Abstract Considerable debate surrounds the question of whether fructosamine concentration should be corrected for serum protein concentration (see 1 for review). Staley (2) has argued against such correction, given that, theoretically, glucose concentration is the rate-limiting step in the glycation reaction; i.e., available lysine residues willalways be in excess of reactive open-chain (carbonyl) glucose molecules, which are only 0.001% of the total (3). However, because the open-chain and cyclic forms of glucose exist in freely exchangeable equilibrium, we conjectured that, as carbonyl glucoses were removed by glycation, more glucose molecules would rapidly isomerize to the open-chainform to maintain the equilibrium, if so, then protein concentration would also be an important factor in determining the glycation rate.

scholarly journals A novel hybrid SEIQR model incorporating the effect of quarantine and lockdown regulations for COVID-19

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
R. Prabakaran ◽  
Sherlyn Jemimah ◽  
Puneet Rawat ◽  
Divya Sharma ◽  
M. Michael Gromiha
Keyword(s):  
Mathematical Models ◽  
Qualitative Agreement ◽  
Mortality Rates ◽  
The Other ◽  
Disease Dynamics ◽  
Disease Propagation ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Geographical Regions ◽  
Rate Limiting

AbstractMitigating the devastating effect of COVID-19 is necessary to control the infectivity and mortality rates. Hence, several strategies such as quarantine of exposed and infected individuals and restricting movement through lockdown of geographical regions have been implemented in most countries. On the other hand, standard SEIR based mathematical models have been developed to understand the disease dynamics of COVID-19, and the proper inclusion of these restrictions is the rate-limiting step for the success of these models. In this work, we have developed a hybrid Susceptible-Exposed-Infected-Quarantined-Removed (SEIQR) model to explore the influence of quarantine and lockdown on disease propagation dynamics. The model is multi-compartmental, and it considers everyday variations in lockdown regulations, testing rate and quarantine individuals. Our model predicts a considerable difference in reported and actual recovered and deceased cases in qualitative agreement with recent reports.

Transport of fluorescein in trichomes of Lycopersicon esculentum

1982 ◽  
Vol 60 (4) ◽  
pp. 397-402 ◽  
Author(s):  
Gregor F. Barclay ◽  
Carol A. Peterson ◽  
Melvin T. Tyree
Keyword(s):  
Lycopersicon Esculentum ◽  
Cytoplasmic Streaming ◽  
Cytochalasin B ◽  
The Other ◽  
Square Root ◽  
Inhibiting Effect ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Other Hand ◽  
Rate Limiting

Translocation of the dye disodium fluorescein (uranin) in trichomes of Lycopersicon esculentum (tomato) was nonpolar and proportional to the square root of time. Inhibition of cytoplasmic streaming by cytochalasin B had no effect on the rate of dye movement. On the other hand, disruption of plasmodesmatal connections between adjacent cells by plasmolysis strongly diminished the rate of fluorescein translocation. Subsequent deplasmolysis of the cells did not remove the inhibiting effect of plasmolysis. The data are consistent with the interpretation that dye movement proceeds by diffusion, the rate-limiting step being transport through plasmodesmatal connections.

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