Antraquinone-triazine derivatives of polysaccharides. Relation between structure and affinity to lactate dehydrogenase

1981 ◽  
Vol 46 (2) ◽  
pp. 419-427 ◽  
Author(s):  
Peter Gemeiner ◽  
Danica Mislovičová ◽  
Jiří Zemek ◽  
Ľudovít Kuniak
Keyword(s):  
Lactate Dehydrogenase ◽  
Degree Of Substitution ◽  
Brilliant Blue ◽  
Rabbit Muscle ◽  
Muscle Lactate ◽  
Cibacron Blue ◽  
Triazine Derivatives ◽  
Covalently Linked ◽  
The Relationship ◽  
Derivatives Of

The inhibition effects (I50) of a series of dyes (Cibacrom Blue 3G-A, Procion Blue MX-R, Remazo Brilliant Blue and Ostazin Brilliant Red S5-B) covalently linked to dextran T 70 were the criterion of the relationship between the structure of antraquinone-triazine compounds and their affinity to rabbit muscle lactate dehydrogenase (LDH). The I50 values indicate a substantial importance of the terminal benzenesulfonane moiety in the structure of Cibacron Blue. Also the degree of substitution of polysaccharide plays an important role in the affinity; its influence upon I50 can be expressed for Cibacron Blue-Dextrans T 70 by an equation of line. The influence of the structure of the polysaccharide constituent on the affinity of Cibacron Blue-(α-glucans) towards LDH is discussed, as well.

Antrachinon-triazine derivatives of polysaccharides, affinity of bovine heart lactate dehydrogenase to Cibacron Blue derivatives of water-soluble dextran fractions and hydroxyethyl-starches

1984 ◽  
Vol 49 (2) ◽  
pp. 549-554 ◽  
Author(s):  
Jana Barthová ◽  
Marie Tichá ◽  
Peter Gemeiner ◽  
Danica Mislovičová
Keyword(s):  
Lactate Dehydrogenase ◽  
Molecular Mass ◽  
Water Soluble ◽  
Relative Molecular Mass ◽  
Cibacron Blue ◽  
Strength Of Interaction ◽  
Triazine Derivatives ◽  
Starch Derivatives ◽  
Derivatives Of ◽  
Hydroxyethyl Starches

The interaction of lactate dehydrogenase with high-molecular-weight derivatives of Cibacron Blue was followed by measuring the reaction kinetics and by means of affinity electrophoresis. The dye was covalently bound to dextrans with relative molecular mass ranging from 5 000 to 2 000 000 and to hydroxyethyl-starches with relative molecular mass from 130 000 to 2 000 000. In the case of dextran derivatives of Cibacron Blue, various degrees of substitution by the dye were tested. Measurement of kinetics showed that the Cibacron Blue derivatives competed with NADH for the binding site on the enzyme and that when the dye was bound to a polysaccharide, the affinity of lactate dehydrogenase to the dye decreased. Both methods used confirmed that the strength of interaction did not depend on the relative molecular mass of the polysaccharide carrier or on the degree of substitution. The interaction of lactate dehydrogenase with hydroxyethyl-starch derivatives of Cibacron Blue was weaker than with dextran derivatives.

scholarly journals Kinetic Studies of Rabbit Muscle Lactate Dehydrogenase

1962 ◽  
Vol 237 (5) ◽  
pp. 1668-1675
Author(s):  
Virginia Zewe ◽  
Herbert J. Fromm
Keyword(s):  
Lactate Dehydrogenase ◽  
Kinetic Studies ◽  
Rabbit Muscle ◽  
Muscle Lactate

scholarly journals Evaluation of equilibrium constants for the interaction of lactate dehydrogenase isoenzymes with reduced nicotinamide-adenine dinucleotide by affinity chromatography

1975 ◽  
Vol 151 (3) ◽  
pp. 631-636 ◽  
Author(s):  
R I Brinkworth ◽  
C J Masters ◽  
D J Winzor
Keyword(s):  
Lactate Dehydrogenase ◽  
Affinity Chromatography ◽  
Equilibrium Constants ◽  
Mouse Tissue ◽  
Rabbit Muscle ◽  
Muscle Lactate ◽  
Sodium Phosphate Buffer ◽  
A Value ◽  
Series Of Experiments ◽  
Reduced Nicotinamide Adenine Dinucleotide

Rabbit muscle lactate dehydrogenase was subjected to frontal affinity chromatography on Sepharose-oxamate in the presence of various concentrations of NADH and sodium phosphate buffer (0.05 M, pH 6.8) containing 0.5 M-NaCl. Quantitative interpretation of the results yields an intrinsic association constant of 9.0 × 104M−1 for the interaction of enzyme with NADH at 5°C, a value that is confirmed by equilibrium-binding measurements. In a second series of experiments, zonal affinity chromatography of a mouse tissue extract under the same conditions was used to evaluate assoication constants of the order 2 × 105M−1, 3 × 105M−1, 4 × 105M−1, 7 × 105M−1 and 2 × 106M−1 for the interaction of NADH with the M4, M3H, M2H2, MH3 and H4 isoenzymes respectively of lactate dehydrogenase.

scholarly journals Covalent labelling of ligand binding sites of human placental S-adenosylhomocysteine hydrolase with 8-azido derivatives of adenosine and cyclic AMP

1985 ◽  
Vol 232 (3) ◽  
pp. 643-650 ◽  
Author(s):  
V N Aiyar ◽  
M S Hershfield
Keyword(s):  
Cyclic Amp ◽  
Binding Sites ◽  
Catalytic Mechanism ◽  
Periodate Oxidation ◽  
Regulatory Subunit ◽  
Adenosylhomocysteine Hydrolase ◽  
Dependent Protein ◽  
Covalently Linked ◽  
The Relationship ◽  
Derivatives Of

S-Adenosylhomocysteine hydrolase (AdoHcyase) has previously been identified as a cytoplasmic adenosine and cyclic AMP binding protein. In order to examine the relationship between the adenosine and cyclic AMP binding sites on this enzyme we have explored the use of 8-azido analogues of adenosine and cyclic AMP as photoaffinity reagents for covalently labelling AdoHcyase purified from human placenta. 8-Azidoadenosine (8-N3-Ado), like adenosine, inactivated AdoHcyase, and the rate of inactivation was greatly increased by periodate oxidation. In addition, 8-N3-Ado was found to participate in the first step in the catalytic mechanism for AdoHcyase, resulting in conversion of enzyme-bound NAD+ to NADH, although it was not a substrate for the full enzyme-catalysed reaction. Radioactively labelled 8-N3-Ado, its periodate-oxidized derivative and 8-azidoadenosine 3′, 5′-phosphate (8-N3-cAMP) bound specifically to adenosine binding sites on AdoHcyase and, after irradiation, became covalently linked to the enzyme. Photoaffinity-labelled enzyme could be precipitated by monoclonal antibody to human AdoHcyase. Two observations suggested that cyclic AMP and adenosine bind to the same sites on AdoHcyase. First cyclic AMP and adenosine each blocked binding of both radioactively labelled 8-N3-Ado and 8-N3-cAMP, and second, digestion with V8 proteinase generated identical patterns of peptides from AdoHcyase that had been photolabelled with [32P]8-N3-cAMP and [3H]8-N3-Ado. Binding sites for cyclic AMP on AdoHcyase were found to differ functionally and structurally from cyclic AMP binding sites on the R1 regulatory subunit of cyclic AMP-dependent protein kinase.

Interactions of Kepone with rabbit muscle lactate dehydrogenase

1978 ◽  
Vol 26 (1) ◽  
pp. 130-133 ◽  
Author(s):  
Bruce M. Anderson ◽  
Susan T. Kohler ◽  
Roderick W. Young
Keyword(s):  
Lactate Dehydrogenase ◽  
Rabbit Muscle ◽  
Muscle Lactate

Inactivation in Vitro of Lactate Dehydrogenase-5 in Blood

1977 ◽  
Vol 14 (1-6) ◽  
pp. 48-52 ◽  
Author(s):  
A. R. Qureshi ◽  
J. H. Wilkinson
Keyword(s):  
Enzyme Activity ◽  
Lactate Dehydrogenase ◽  
Whole Blood ◽  
Rabbit Muscle ◽  
Muscle Lactate ◽  
Rabbit Blood

During incubation with rabbit blood in vitro rabbit-muscle lactate dehydrogenase-5 was inactivated at a rate similar to that observed in vivo. By contrast plasma and plasma containing erythrocytes had no effect on the enzyme activity, but plasma containing leucocytes inactivated the enzyme at the same rate as whole blood. The results obtained support the concept that intravascular inactivation accounts for the disappearance of enzymes from the circulation.

scholarly journals The mechanism of adduct formation between NAD+ and pyruvate bound to pig heart lactate dehydrogenase

1979 ◽  
Vol 177 (3) ◽  
pp. 951-957 ◽  
Author(s):  
D C Wilton
Keyword(s):  
Lactate Dehydrogenase ◽  
Dissociation Constant ◽  
Active Site ◽  
Adduct Formation ◽  
Rabbit Muscle ◽  
Muscle Lactate

1. The rate of adduct formation between NAD+ and enol-pyruvate at the active site of lactate dehydrogenase is determined by the rate of enolization of pyruvate in solution. 2. The proportion of enol-pyruvate solutions is less than 0.01%. 3. The overall dissociation constant of adduct formation is less than 5 × 10(-8) M for pig heart lactate dehydrogenase at pH 7.0. 4. The unusual kinetics for adduct formation previously observed in the case of rabbit muscle lactate dehydrogenase [Griffin & Criddle (1970) Biochemistry 9, 1195–1205] may be attributed to the concentration of enol-pyruvate in solution being considerably less than the concentration of enzyme.

Flow kinetics of lactate dehydrogenase chemically attached to nylon tubing

1978 ◽  
Vol 56 (8) ◽  
pp. 774-779 ◽  
Author(s):  
N. J. Daka ◽  
K. J. Laidler
Keyword(s):  
Lactate Dehydrogenase ◽  
Flow Rate ◽  
Rabbit Muscle ◽  
Modified Technique ◽  
Muscle Lactate ◽  
Flow Rates ◽  
Diffusion Controlled ◽  
Michaelis Constants ◽  
Reduced Nicotinamide Adenine Dinucleotide ◽  
Nylon Tubing

Rabbit muscle lactate dehydrogenase (EC 1.1.1.27) was attached covalently to the inner surface of nylon tubing; a modified technique, involving benzidine and glutaraldehyde, was used, and the resulting immobilized enzyme showed no loss of activity over a period of several months. An experimental study was made of the flow kinetics for the reaction between pyruvate and reduced nicotinamide adenine dinucleotide in two limiting cases, one substrate in excess and the concentration of the other one varied. A range of flow rates and temperatures was covered. The results were analyzed in various ways on the basis of the Kobayashi–Laidler treatment of flow systems. It was concluded that the kinetics are largely diffusion-controlled, especially at the lower substrate concentrations and flow rates. The values of the apparent Michaelis constants vary with flow rate vf, being linear in vf−1/3, and the values extrapolated to infinite flow rate (vf−1/3 = 0) approach the values for the enzyme in free solution. Analysis of the rates led to activation energies for the diffusion of the two substrates.

Sign in / Sign up

Export Citation Format

Share Document