scholarly journals The reaction of nitrite with the haemocyanin of Astacus leptodactylus

1988 ◽  
Vol 249 (3) ◽  
pp. 891-896 ◽  
Author(s):  
J P Tahon ◽  
D Van Hoof ◽  
C Vinckier ◽  
R Witters ◽  
M De Ley ◽  
...  
Keyword(s):  
Active Site ◽  
Active Sites ◽  
Reaction Rate ◽  
Anaerobic Treatment ◽  
Astacus Leptodactylus ◽  
First Order ◽  
Bridging Ligand ◽  
A Value ◽  
Pseudo First Order

The reaction of nitrite at pH 5.7 with deoxyhaemocyanin of Astacus leptodactylus yielded methaemocyanin in two one-electron steps, as nitrite was reduced to NO. This methaemocyanin could be almost fully regenerated by an anaerobic treatment with HONH2, in contrast with the methaemocyanin prepared with H2O2. A destruction of active sites on treating oxyhaemocyanin with HONH2 explains the partial regeneration of methaemocyanin under air, as traces of H2O2 are formed in the autoxidation of HONH2. The reaction rate of nitrite with deoxyhaemocyanin is almost 15 times that with oxyhaemocyanin. The slope of -1.0 for the logarithm of the pseudo-first-order rate constants plotted against pH indicates that HNO2 is the reacting species. Methaemocyanin was e.p.r.-undetectable, but a binuclear signal was observed at g = 2 on binding nitrite to methaemocyanin. This signal disappeared with a pKa of 6.50, suggesting that a mu-aquo bridging ligand, which can be replaced by nitrite, is deprotonated to a mu-hydroxo bridging ligand, which resists substitution by nitrite. The intensity of this triplet e.p.r. signal allowed the determination of the association constant of nitrite to the active site of Astacus methaemocyanin and yielded a value of 237 M-1 at pH 5.7. The interpretation by some authors of nitrosylhaemocyanin as a nitrite derivative of semimethaemocyanin is contradicted by this rapid reaction of nitrite with copper(I) in deoxyhaemocyanin and in semi-methaemocyanin and by the low binding constant of nitrite to the active site of methaemocyanin.

scholarly journals The reaction of nitrite with the haemocyanin of the Roman snail (Helix pomatia)

1990 ◽  
Vol 271 (3) ◽  
pp. 779-783 ◽  
Author(s):  
J P Tahon ◽  
G Maes ◽  
C Vinckier ◽  
R Witters ◽  
T Zeegers-Huyskens ◽  
...  
Keyword(s):  
Active Site ◽  
Reaction Rate ◽  
Helix Pomatia ◽  
Astacus Leptodactylus ◽  
Raman Spectrometry ◽  
Signal Characteristic ◽  
Bridging Ligand ◽  
Pka Value ◽  
Snail Helix Pomatia ◽  
Hasselbalch Equation

The reaction of nitrite at pH 5.0-7.0 with the deoxyhaemocyanin of a mollusc, the Roman snail (Helix pomatia), yielded nitrosylhaemocyanin (CuIA.NO+ CuIIB), in contrast with the formation of methaemocyanin with the deoxyhaemocyanin of the crustacean Astacus leptodactylus (mud crayfish). With Helix haemocyanin 1 NO was thereby liberated per active site, as shown by m.s., as against 2 NO with Astacus haemocyanin. Helix nitrosylhaemocyanin was characterized in c.d. by the negative extremum at 336 nm (CuIA.NO+) and by the mononuclear e.p.r. signal at g = 2 (CuIIB). Binuclear e.p.r. signals have been observed after the addition of nitrite to methaemocyanins. With Astacus methaemocyanin, no further reaction occurred, whereas with Helix methaemocyanin the mononuclear e.p.r. signal, characteristic for nitrosylhaemocyanin gradually appeared. This formation of Helix nitrosylhaemocyanin implicates the binding, most likely on CuIIA, of a second nitrite besides a bridging nitrite, so that a dismutation into NO and NO2 can occur there. A further dismutation of NO2 yields nitrite and nitrate. The formation of the latter was demonstrated by Raman spectrometry. The reaction rate of Helix methaemocyanin with nitrite decreased with increasing pH according to the Henderson-Hasselbalch equation with a pKa value of 6.77, attributed to a mu-aquo bridging ligand, which can be exchanged for nitrite, in equilibrium with a mu-hydroxo ligand which cannot. These data also favour the formulation of the final reaction product as nitrosylhaemocyanin instead of semi-methaemocyanin, with or without bound nitrite.

Nonlinear Vibrations in a 2-Dimensional Protein Cluster Model with Linear Bonds

2000 ◽  
Vol 214 (1) ◽  
Author(s):  
E.G. Shidlovskaya ◽  
L. Schimansky-Geier ◽  
Yu.M. Romanovsky
Keyword(s):  
Active Site ◽  
Internal Resonance ◽  
Cluster Model ◽  
Active Sites ◽  
Nonlinear Vibrations ◽  
Nonlinear Phenomena ◽  
Two Dimensional ◽  
Dimensional Model ◽  
Two Dimensional Model

A two dimensional model for the substrate inside a pocket of an active site of an enzyme is presented and investigated as a vibrational system. The parameters of the system are evaluated for α-chymotrypsin. In the case of internal resonance it is analytically and numerically shown that the energy concentrated on a certain degree of freedom might be several times larger than in the non-resonant case. Additionally, the system is driven by harmonic excitations and again energy due to nonlinear phenomena is redistributed inhomogeneously. These results may be of importance for the determination of the rates of catalytic events of substrates bound in pockets of active sites.

Direct spectrophotometry of magnesium in serum after reaction with hexokinase and glucose-6-phosphate dehydrogenase.

1985 ◽  
Vol 31 (5) ◽  
pp. 703-705 ◽  
Author(s):  
M Tabata ◽  
T Kido ◽  
M Totani ◽  
T Murachi
Keyword(s):  
Spectrophotometric Method ◽  
Present Method ◽  
Reaction Rate ◽  
Colorimetric Method ◽  
Simple Method ◽  
True Value ◽  
A Value ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Direct Spectrophotometry

Abstract We describe a simple method for determining magnesium in serum by using hexokinase (EC 2.7.1.1) and glucose-6-phosphate dehydrogenase (EC 1.1.1.49). The method is based on determination of the reaction rate of hexokinase activated by Mg2+, which participates in the hexokinase reaction as the substrate in the form of a Mg X ATP2- complex. The reaction rate is determined from the change in absorbance at 340 nm as NADPH is produced by the action of glucose-6-phosphate dehydrogenase. This simple and rapid spectrophotometric method does not require expensive instrumentation, but results correlate satisfactorily with those obtained by atomic absorption spectroscopy. Thus, the present method gives a "true" value for magnesium in serum, a value appreciably lower than that obtained by an earlier colorimetric method, the Xylidyl Blue II method (Biochem Med 7: 208-217, 1973), which lacks specificity.

scholarly journals Acrolein, an irreversible active-site-directed inhibitor of deoxyribose 5-phosphate aldolase?

1976 ◽  
Vol 153 (2) ◽  
pp. 495-497 ◽  
Author(s):  
D C Wilton
Keyword(s):  
Schiff Base ◽  
Rate Constant ◽  
Active Site ◽  
Order Rate Constant ◽  
Lysine Residue ◽  
Prolonged Incubation ◽  
Enzyme Active Site ◽  
First Order ◽  
Substrate Analogue ◽  
Pseudo First Order

The enzyme deoxyribose 5-phosphate aldolase was irreversibly inactivated by the substrate analogue acrolein with a pseudo-first-order rate constant of 0.324 min-1 and a Ki (apparent) of 2.7 × 10(-4) m. No inactivation was observed after prolonged incubation with the epoxide analogues glycidol phosphate and glycidaldehyde. It is suggested that the acrolein is first activated by forming a Schiff base with the enzyme active-site lysine residue and it is the activated inhibitor that reacts with a suitable-active-site nucleophile.

Kinetics of the Ozonation of Tetrabromobisphenol-A in Wastewater

2011 ◽  
Vol 383-390 ◽  
pp. 2945-2950 ◽  
Author(s):  
Jie Zhang ◽  
Shi Long He ◽  
Mei Feng Hou ◽  
Li Ping Wang ◽  
Li Jiang Tian
Keyword(s):  
Rate Constants ◽  
Apparent Rate Constant ◽  
Reaction Rate ◽  
Batch Reactor ◽  
Kinetic Studies ◽  
Tetrabromobisphenol A ◽  
First Order ◽  
Reaction Rate Constants ◽  
Pseudo First Order ◽  
Kinetics Of

The kinetics of TBBPA degradation by ozonation in semi-batch reactor was studied. The reaction rate constants of TBBPA with O3 and •OH were measured by means of direct ozone attack and competition kinetics, and the values of which were 6.10 l/(mol•s), 4.8×109 l/(mol•s), respectively. Results of kinetic studies showed that TBBPA degradation by ozonation under the different conditions tested followed the pseudo-first-order. The values of apparent rate constant of TBBPA degradation increased with the increase of ozone dosage and pH, but decreased with the increase of initial TBBPA concentration.

scholarly journals Probing the active site residues in aromatic donor oxidation in horseradish peroxidase: involvement of an arginine and a tyrosine residue in aromatic donor binding

1996 ◽  
Vol 314 (3) ◽  
pp. 985-991 ◽  
Author(s):  
Subrata ADAK ◽  
Abhijit MAZUMDER ◽  
Ranajit K. BANERJEE
Keyword(s):  
Active Site ◽  
Native Enzyme ◽  
Tyrosine Residue ◽  
Tyrosine Residues ◽  
First Order ◽  
First Order Kinetics ◽  
Aromatic Donor ◽  
Pseudo First Order ◽  
Modified Enzyme ◽  
Compound Ii

The plausible role of arginine and tyrosine residues at the active site of horseradish peroxidase (HRP) in aromatic donor (guaiacol) oxidation was probed by chemical modification followed by characterization of the modified enzyme. The arginine-specific reagents phenylglyoxal (PGO), 2,3-butanedione and 1,2-cyclohexanedione all inactivated the enzyme, following pseudo-first-order kinetics with second-order rate constants of 24 M-1·min-1, 0.8 M-1·min-1 and 0.54 M-1·min-1 respectively. Modification with tetranitromethane, a tyrosine-specific reagent, also resulted in 50% loss of activity following pseudo-first-order kinetics with a second-order rate constant of 2.0 M-1·min-1. The substrate, H2O2, and electron donors such as I- and SCN- offered no protection against inactivation by both types of modifier, whereas the enzyme was completely protected by guaiacol or o-dianisidine, an aromatic electron donor (second substrate) oxidized by the enzyme. These studies indicate the involvement of arginine and tyrosine residues at the aromatic donor site of HRP. The guaiacol-protected phenylglyoxal-modified enzyme showed almost the same binding parameter (Kd) as the native enzyme, and a similar free energy change (∆G´) for the binding of the donor. Stoicheiometric studies with [7-14C]phenylglyoxal showed incorporation of 2 mol of phenylglyoxal per mol of enzyme, indicating modification of one arginine residue for complete inactivation. The difference absorption spectrum of the tetranitromethane-modified against the native enzyme showed a peak at 428 nm, characteristic of the nitrotyrosyl residue, that was abolished by treatment with sodium dithionite, indicating specific modification of a tyrosine residue. Inactivation stoicheiometry showed that modification of one tyrosine residue per enzyme caused 50% inactivation. Binding studies by optical difference spectroscopy indicated that the arginine-modified enzyme could not bind guaiacol at all, whereas the tyrosine-modified enzyme bound it with reduced affinity (Kd 35 mM compared with 10 mM for the native enzyme). Both the modified enzymes, however, retained the property of the formation of compound II (one-electron oxidation state higher than native ferriperoxidase) with H2O2, but reduction of compound II to native enzyme by guaiacol did not occur in the PGO-modified enzyme, owing to lack of binding. No non-specific change in protein structure due to modification was evident from circular dichroism studies. We therefore suggest that the active site of HRP for aromatic donor oxidation is composed of an arginine and an adjacent tyrosine residue, of which the former plays an obligatory role in aromatic donor binding whereas the latter residue plays a facilitatory role, presumably by hydrophobic interaction or hydrogen bonding.

Enhancement of decomposition of 2-chlorophenol with ultrasound/H2O2 process

1996 ◽  
Vol 34 (9) ◽  
pp. 41-48 ◽  
Author(s):  
Jih-Gaw Lin ◽  
Cheng-Nan Chang ◽  
Jer-Ren Wu ◽  
Ying-Shih Ma
Keyword(s):  
Ionic Strength ◽  
Reaction Rate ◽  
Order Reaction ◽  
Pseudo First Order Reaction ◽  
Initial Concentration ◽  
First Order ◽  
Toc Removal ◽  
Cp Decomposition ◽  
Effects Of Ph ◽  
Pseudo First Order

We investigated the effects of pH, ionic strength, catalyst, and initial concentration on both decomposition of 2-chlorophenol (2-cp) and removal of total organic carbon (TOC) in aqueous solution with ultrasonic amplitude 120 μm and H2O2 (200 mg/l). When the initial concentrations of 2-cp was 100 mg/l and the pH was controlled at 3, the rate of 2-cp decomposition was enhanced up to 6.6-fold and TOC removal up to 9.8-fold over pH controlled at 11. At pH 3, the efficiency of decomposition of 2-cp was 99% but the removal of TOC was only 63%; a similar situation applied at pH 7 and 11. Hence intermediate compounds were produced and 2-cp was not completely mineralized. When the concentration of ionic strength was increased from 0.001 to 0.1 M, the rate of 2-cp decomposition was enhanced only 0.3-fold, whereas the TOC removal was not enhanced. In comparison of the effects of pH and ionic strength, pH had greater influence on both 2-cp decomposition and TOC removal than ionic strength. The effect of a catalyst (FeSO4) on decomposition of 2-cp was insignificant comparing with direct addition of H2O2. The reaction rate at a smaller initial concentration of 2-cp (10 mg/l) was more rapid than at a greater one (100 mg/l). The rate of 2-cp decomposition and TOC removal appeared to follow pseudo-first-order reaction kinetics.

scholarly journals Determination of the Bridging Ligand in the Active Site of Tyrosinase

Molecules ◽  
2017 ◽  
Vol 22 (11) ◽  
pp. 1836 ◽  
Author(s):  
Congming Zou ◽  
Wei Huang ◽  
Gaokun Zhao ◽  
Xiao Wan ◽  
Xiaodong Hu ◽  
...  
Keyword(s):  
Active Site ◽  
Bridging Ligand

scholarly journals Effect of precursor type on the reduction of concentrated nitrate using zero-valent copper and sodium borohydride

2017 ◽  
Vol 77 (1) ◽  
pp. 114-122 ◽  
Author(s):  
Tihitinna Asmellash Belay ◽  
C. Y. Lin ◽  
H. M. Hsiao ◽  
M. F. Chang ◽  
J. C. Liu
Keyword(s):  
Particle Size ◽  
Surface Area ◽  
Sodium Borohydride ◽  
Reaction Rate ◽  
Reduction Rate ◽  
First Order ◽  
Reducing Conditions ◽  
Pseudo First Order

Abstract In this study, we demonstrated that the choice of precursor has a strong effect on the reduction of nitrate (NO3−) using zero-valent copper (Cu0) synthesized by sodium borohydride (NaBH4). Different precursors: CuSO4, CuO, Cu2O, Cu powder, and Cu mesh were used to reduce NO3− at 677 mg-N/L under the reducing conditions of NaBH4. Compared with the prehydrolyzed samples, those prepared without prehydrolysis exhibited lower reduction rates, longer times and higher concentrations of nitrite (NO2−) intermediate. It was found that one-time addition of NaBH4 resulted in higher reduction rate and less NO2− intermediate than two-step addition. Results showed that Cu0 from CuSO4 possessed the smallest particle size (890.9 nm), highest surface area (26.0 m2/g), and highest reaction rate (0.166 min−1). Values of pseudo-first-order constant (kobs) were in the order: CuSO4 > CuO > Cu2O > Cu powder >Cu mesh. However, values of surface area-normalized reaction rate (kSA) were approximately equal. It was proposed that NO3− was reduced to NO2− on Cu0, and then converted to NH4+ and N2, respectively; H2 generated from both NaBH4 hydration and Cu (II) reduction contributed to NO3− reduction as well.

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