scholarly journals The stereochemical course of phosphoryl transfer catalysed by glucokinase.

1982 ◽  
Vol 201 (2) ◽  
pp. 421-423 ◽  
Author(s):  
D Pollard-Knight ◽  
B V L Potter ◽  
P M Cullis ◽  
G Lowe ◽  
A Cornish-Bowden
Keyword(s):  
Rat Liver ◽  
Rate Constants ◽  
Ternary Complex ◽  
The Other ◽  
Chemical Mechanism ◽  
Phosphoryl Group ◽  
Phosphoryl Transfer ◽  
Reaction Proceeds ◽  
Gamma S ◽  
Yeast Hexokinase

Adenosine 5'-[gamma(S)-16O,17O,18O]triphosphate has been used to determine the stereo-chemical course of phosphoryl transfer catalysed by rat liver glucokinase. The chirality of the product, D-glucose 6-[16O,17O,18O]phosphate was analysed by 31P n.m.r. spectroscopy. The reaction proceeds with inversion of configuration at phosphorus. The simplest interpretation of this result, which is the same as that observed with yeast hexokinase [Lowe & Potter (1981) Biochem. J. 199, 277-233], is that the phosphoryl group is transferred between MgATP2- and glucose in the ternary complex by an ‘in-line’ mechanism. It accords with the veiw that the kinetic differences between glucokinase and the other hexokinases arise from differences in rate constants and not from any fundamental differences in chemical mechanism.

scholarly journals The stereochemical course of yeast hexokinase-catalysed phosphoryl transfer by using adenosine 5′[γ(S)-16O,17O,18O]triphosphate as substrate

1981 ◽  
Vol 199 (1) ◽  
pp. 227-233 ◽  
Author(s):  
G Lowe ◽  
B V L Potter
Keyword(s):  
Ternary Complex ◽  
Phosphoryl Group ◽  
Phosphoryl Transfer ◽  
Enzyme Substrate ◽  
Gamma S ◽  
Yeast Hexokinase

Adenosine 5′[gamma(S)-16O, 17O, 18O]triphosphate has been synthesized and used to determine the stereochemical course of phosphoryl transfer catalysed by yeast hexokinase. The chirality at phosphorus of the D-glucose 6-[16O,17O,18O]phosphate formed was analysed, after cyclization and methylation, by 31P n.m.r. spectroscopy. The phosphoryl transfer was found to occur with inversion of configuration, with a stereoselectivity in excess of 94%. The simplest interpretation of this result is that the phosphoryl group is transferred between substrates in the enzyme-substrate ternary complex by an ‘in line’ mechanism.

KINETICS FOR THE INHIBITION OF CARBOXYLESTERASE BY MALAOXON

1967 ◽  
Vol 45 (6) ◽  
pp. 757-771 ◽  
Author(s):  
A. R. Main ◽  
W. C. Dauterman
Keyword(s):  
Rat Liver ◽  
Rate Constants ◽  
Direct Evidence ◽  
The Other ◽  
Velocity Constant ◽  
Irreversible Inhibition ◽  
Inhibition Reaction ◽  
Phosphorylation Rate

Malaoxon and carboxylesterase undergo two separate but simultaneous reactions when mixed in solution. One results in the irreversible inhibition of carboxylesterase. In the other, malaoxon acts as a substrate and is hydrolyzed.This work is concerned primarily with the inhibition reaction, although direct evidence of the substrate reaction is also given. It seemed possible that inhibition could occur by two mutually exclusive routes. Equations were derived for each of the two routes, and both equations gave identical and unambiguous expressions for ki, the bimolecular velocity constant of the inhibition reaction. A procedure for determining the kivalues of malaoxon and of four carboalkoxy homologues of malaoxon in reaction with partially purified rat-liver carboxylesterase is described. Evidence is given which suggested that the substrate reaction was controlled by the acylation step. This in turn suggested that useful estimates of the binding (Ka) and phosphorylation rate constants (k2p) could be made. Values for kiand tentative values for k2pand Kaare given.

scholarly journals The stereochemical course of phosphoryl transfer catalysed by polynucleotide kinase (bacteriophage-T4-infected Escherichia coli B)

1981 ◽  
Vol 199 (1) ◽  
pp. 273-276 ◽  
Author(s):  
R L Jarvest ◽  
G Lowe
Keyword(s):  
Escherichia Coli ◽  
Phosphorus Atom ◽  
Bacteriophage T4 ◽  
Phosphoryl Group ◽  
Phosphoryl Transfer ◽  
Polynucleotide Kinase ◽  
Gamma S ◽  
Escherichia Coli B

Polynucleotide kinase (bacteriophage-T4-infected Escherichia coli B) catalyses the transfer of the [gamma-16O,17O,18O]phosphoryl group from 5′[gamma(S)-16O,17O,18O]ATP to 3′-AMP with inversion of configuration at the phosphorus atom. The simplest interpretation of this observation is that the [gamma-16O,17O,18O]phosphoryl group is transferred directly from ATP to the co-substrate by an ‘in-line’ mechanism.

Purification and kinetic characterization of hexokinase and glucose-6-phosphate dehydrogenase fromSchizosaccharomyces pombe

1998 ◽  
Vol 76 (1) ◽  
pp. 107-113 ◽  
Author(s):  
C Stan Tsai ◽  
Q Chen
Keyword(s):  
Pentose Phosphate Pathway ◽  
Ternary Complex ◽  
Kinetic Studies ◽  
Product Inhibition ◽  
Pentose Phosphate ◽  
Oxidative Pentose Phosphate Pathway ◽  
Phosphoryl Group ◽  
Maximal Velocity ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Yeast Hexokinase

Hexokinase and D-glucose-6-phosphate dehydrogenase (G6PDH) from Schizosaccharomyces pombe have been purified 250-fold by an identical three-step. Both enzymes are dimeric with a molecular mass of 88 kDa for the kinase and 112 kDa for the dehydrogenase. Steady-state kinetic studies were performed on hexokinase and G6PDH, which form the glucose phosphate branch of the oxidative pentose phosphate pathway of S. pombe (fission yeast). Hexokinase promotes Mg2+-activated phosphorylation of D-glucose by the equilibrium random Bi Bi mechanism with formation of the abortive enzyme-ADP-glucose complex. ADP inhibits the kinase competitively versus ATP and noncompetitively versus D-glucose. The Mg2+activation of hexokinase is associated with an increase in the maximal velocity by its interaction with the ternary complex to facilitate the transfer of the phosphoryl group. G6PDH catalyzes NADP+-linked oxidation of D-glucose-6-phosphate by the ordered Bi Bi mechanism with NADP+as the leading reactant. High NADP+concentration inhibits the dehydrogenase by forming the dead-end ternary complex. In addition, G6PDH is also subjected to product inhibition by NADPH and noncompetitive inhibition by A(G)TP. Thus, the oxidative pentose phosphate pathway in S. pombe may be regulated via inhibition of hexokinase by ADP in conjunction with inhibition of G6PDH by NADPH and ATP.Key words: yeast hexokinase, glucose-6-phosphate dehydrogenase.

Methylene. III. Gas phase reactions with 1-chloropropane

1969 ◽  
Vol 312 (1509) ◽  
pp. 141-161 ◽  
Keyword(s):  
Gas Phase ◽  
Rate Constants ◽  
Hydrogen Abstraction ◽  
The Other ◽  
Gas Phase Reactions ◽  
Other Hand ◽  
Chlorine Substituent ◽  
High Degree ◽  
Singlet Methylene

The work described in this and the following paper is a continuation of that in parts I and II, devoted to elucidation of the mechanism of the reactions of methylene with chloroalkanes, with particular reference to the reactivities of singlet and triplet methylene in abstraction and insertion processes. The products of the reaction between methylene, prepared by the photolysis of ketene, and 1-chloropropane have been identified and estimated and their dependence on reactant pressures, photolysing wavelength and presence of foreign gases (oxygen and carbon mon­oxide) has been investigated. Both insertion and abstraction mechanisms contribute significantly to the over-all reaction, insertion being relatively much more important than with chloroethane. This type of process appears to be confined to singlet methylene. If, as seems likely, there is no insertion into C—Cl bonds under our conditions (see part IV), insertion into C2—H and C3—H bonds occurs in statistical ratio, approximately. On the other hand, the chlorine substituent reduces the probability of insertion into C—H bonds in its vicinity. As in the chloroethane system, both species of methylene show a high degree of selectivity in their abstraction reactions. We find that k S Cl / k S H >7.7, k T Cl / k T H < 0.14, where the k ’s are rate constants for abstraction, and the super- and subscripts indicate the species of methylene and the type of atom abstracted, respectively. Triplet methylene is discriminating in hydrogen abstraction from 1-C 3 H 7 Cl, the overall rates for atoms attached to C1, C2, C3 being in the ratios 2.63:1:0.

scholarly journals The fractionation of nuclei from mammalian cells by zonal centrifugation

1968 ◽  
Vol 109 (1) ◽  
pp. 127-135 ◽  
Author(s):  
I R Johnston ◽  
A P Mathias ◽  
F. Pennington ◽  
D. Ridge
Keyword(s):  
Rat Liver ◽  
Mammalian Cells ◽  
Mouse Liver ◽  
The Other ◽  
Slow Speed ◽  
Adult Rats ◽  
Sucrose Solutions ◽  
Liver Nuclei ◽  
Effect Of Age ◽  
Zonal Rotor

1. Purified liver nuclei from adult rats separate into two main zones when centrifuged in the slow-speed zonal rotor. One zone contains diploid nuclei, the other tetraploid. 2. The effect of age on the pattern of rat liver ploidy was examined. Tetraploid nuclei are virtually absent from young animals. They increase in proportion steadily with age. Partial hepatectomy disturbs the pattern of ploidy. 3. The zonal centrifuge permits the separation of diploid, tetraploid, octaploid and hexadecaploid nuclei from mouse liver. 4. Rat liver nuclei are isopycnic with sucrose solutions of density 1·35 at 5°.

scholarly journals DABCO Catalyzed Synthesis of Xanthene Derivatives in Aqueous Media

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Pradeep Paliwal ◽  
Srinivasa Rao Jetti ◽  
Anjna Bhatewara ◽  
Tanuja Kadre ◽  
Shubha Jain
Keyword(s):  
Reaction Time ◽  
Aqueous Media ◽  
Environmentally Friendly ◽  
The Other ◽  
Short Reaction Time ◽  
Xanthene Derivatives ◽  
Reaction Proceeds ◽  
Conventional Methods

The reaction of 5,5-dimethylcyclohexane-1,3-dione with various heteroarylaldehydes afforded the corresponding heteroaryl substituted xanthene derivatives 1(a–f). The reaction proceeds via the initial Knoevenagel, subsequent Michael, and final heterocyclization reactions using 1,4-diazabicyclo[2.2.2]octane (DABCO) as a catalyst in aqueous media. The synthesized heteroaryl substituted xanthenes 1(a–f) reacted with malononitrile to obtain different alkylidenes 2(a–f). Short reaction time, environmentally friendly procedure, avoiding of cumbersome apparatus, and excellent yields are the main advantages of this procedure which makes it more economic than the other conventional methods.

scholarly journals Ganglioside incorporation and release by the isolated perfused rat liver

1991 ◽  
Vol 274 (2) ◽  
pp. 581-585 ◽  
Author(s):  
S C Kivatinitz ◽  
A Miglio ◽  
R Ghidoni
Keyword(s):  
Rat Liver ◽  
The Other ◽  
Regulatory Role ◽  
Isolated Perfused Rat Liver ◽  
Order Kinetic ◽  
Perfused Rat Liver ◽  
Ganglioside Gm1 ◽  
First Order ◽  
Pseudo First Order

The fate of exogenous ganglioside GM1 labelled in the sphingosine moiety, [Sph-3H]GM1, administered as a pulse, in the isolated perfused rat liver was investigated. When a non-recirculating protocol was employed, the amount of radioactivity in the liver and perfusates was found to be dependent on the presence of BSA in the perfusion liquid and on the time elapsed after the administration of the ganglioside. When BSA was added to the perfusion liquid, less radioactivity was found in the liver and more in the perfusate at each time tested, for up to 1 h. The recovery of radioactivity in the perfusates followed a complex course which can be described by three pseudo-first-order kinetic constants. The constants, in order of decreasing velocity, are interpreted as: (a) the dilution of the labelled GM1 by the constant influx of perfusion liquid; (b) the washing off of GM1 loosely bound to the surface of liver cells; (c) the release of gangliosides from the liver. Process (b) was found to be faster in the presence of BSA, probably owing to the ability of BSA to bind gangliosides. The [Sph-3H]GM1 in the liver underwent metabolism, leading to the appearance of products of anabolic (GD1a, GD1b) and catabolic (GM2, GM3) origin; GD1a appeared before GM2 and GM3 but, at times longer than 10 min, GM2 and GM3 showed more radioactivity than GD1a. At a given time the distribution of the radioactivity in the perfusates was quite different from that of the liver. In fact, after 60 min GD1a was the only metabolite present in any amount, the other being GM3, the quantity of which was small. This indicates that the liver is able to release newly synthesized gangliosides quite specifically. When a recirculating protocol was used, there were more catabolites and less GD1a than with the non-recirculating protocol. A possible regulatory role of ganglioside re-internalization on their own metabolism in the liver is postulated.

scholarly journals The thermal decomposition of nitrogen pentoxide at low pressures

1925 ◽  
Vol 109 (751) ◽  
pp. 526-540 ◽  
Keyword(s):  
Thermal Decomposition ◽  
The Other ◽  
Nitrogen Tetroxide ◽  
Nitrogen Pentoxide ◽  
Other Hand ◽  
Rate Of Reaction ◽  
Reaction Proceeds ◽  
Products Of Reaction ◽  
Glass Surfaces ◽  
Low Pressures

Unimolecular reactions possess a unique interest in that, as Perrin (‘Ann. Physique,’ vol. 11, p. 5, 1919) first pointed out, for the occurrence of such, some type of interaction between radiation and matter must take place. Although such reactions appear to be extremely rare, many physical processes such as evaporation, ionisation in gases at high temperatures and radio-active decay, proceed at rates conforming to a unimolecular law; true chemical reactions which are definitely unimolecular and not pseudo-unimolecular in character are, on the other hand, stated by many ( e. g ., Lowry, ‘Trans. Farad. Soc.,’ vol. 17, p. 596 (1922) ) to be non-existent. In order to substantiate this statement, it is clearly necessary to prove the more complex character of any reaction which satisfies the usual criteria of unimolecular change. The thermal decomposition of gaseous nitrogen pentoxide apparently fulfils these conditions, for Daniels and Johnston (‘J. Am. C. S.,’ vol. 43, p. 53 (1921)) showed that the reaction proceeded according to a unimolecular law over wide ranges of variation of pressure, and Lueck ( ibid ., vol. 44, p. 757 (1922)) obtained practically identical unimolecular constants for the decomposition in solution in carbon tetrachloride and chloroform. On the other hand, Daniels, Wulf and Karrer ( ibid ., vol. 44, p. 2402 (1922) ) suspected the reaction to be autocatalytic, owing to the apparent retardation of the reaction velocity in the presence of ozone, but the experiments of one of us (Hirst, ‘J. C. S.,’ vol. 127, p. 657 (1925), and of White and Tolman (‘J. Am. C. S.’ vol. 47, p. 1,240 (1925)) proved this to be erroneous. In addition, it has been shown that the reaction proceeds uniformly according to the unimolecular law even in the presence of extensive glass surfaces, or of gases which may be either indifferent, such as argon and nitrogen, or the products of reaction, such as nitrogen tetroxide or dioxide or oxygen. The rate of reaction may be expressed in the form - d C/ dt = 4·98 × 10 13 e -24.700/RT . C. Attempts have been made to interpret the experimental results on the hypothesis that the reaction is in reality bimolecular, and only apparently unimolecular in character; but owing to the abnormally large value of the energy of activation, namely, 24,700 calories per gram. molecule, the number of molecules which could be activated per second by inelastic collision, calculated according to the kinetic theory, falls far short of the observed reaction rate, being, in fact, some 10 5 times smaller.

scholarly journals Subcellular distribution and characteristics of trihydroxycoprostanoyl-CoA synthetase in rat liver

1989 ◽  
Vol 257 (1) ◽  
pp. 221-229 ◽  
Author(s):  
L Schepers ◽  
M Casteels ◽  
K Verheyden ◽  
G Parmentier ◽  
S Asselberghs ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Cholic Acid ◽  
Rat Liver ◽  
Subcellular Distribution ◽  
The Other ◽  
Acid Activation ◽  
Long Chain Fatty Acids ◽  
Microsomal Membranes ◽  
Triton X ◽  
Long Chain

The subcellular distribution and characteristics of trihydroxycoprostanoyl-CoA synthetase were studied in rat liver and were compared with those of palmitoyl-CoA synthetase and choloyl-CoA synthetase. Trihydroxycoprostanoyl-CoA synthetase and choloyl-CoA synthetase were localized almost completely in the endoplasmic reticulum. A quantitatively insignificant part of trihydroxycoprostanoyl-CoA synthetase was perhaps present in mitochondria. Peroxisomes, which convert trihydroxycoprostanoyl-CoA into choloyl-CoA, were devoid of trihydroxycoprostanoyl-CoA synthetase. As already known, palmitoyl-CoA synthetase was distributed among mitochondria, peroxisomes and endoplasmic reticulum. Substrate- and cofactor- (ATP, CoASH) dependence of the three synthesis activities were also studied. Cholic acid and trihydroxycoprostanic acid did not inhibit palmitoyl-CoA synthetase; palmitate inhibited the other synthetases non-competitively. Likewise, cholic acid inhibited trihydroxycoprostanic acid activation non-competitively and vice versa. The pH curves of the synthetases did not coincide. Triton X-100 affected the activity of each of the synthetases differently. Trihydroxycoprostanoyl-CoA synthetase was less sensitive towards inhibition by pyrophosphate than choloyl-CoA synthetase. The synthetases could not be solubilized from microsomal membranes by treatment with 1 M-NaCl, but could be solubilized with Triton X-100 or Triton X-100 plus NaCl. The detergent-solubilized trihydroxycoprostanoyl-CoA synthetase could be separated from the solubilized choloyl-CoA synthetase and palmitoyl-CoA synthetase by affinity chromatograpy on Sepharose to which trihydroxycoprostanic acid was bound. Choloyl-CoA synthetase and trihydroxycoprostanoyl-CoA synthetase could not be detected in homogenates from kidney or intestinal mucosa. The results indicate that long-chain fatty acids, cholic acid and trihydroxycoprostanic acid are activated by three separate enzymes.

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