Topic:Enzymes
Enzymes are biological catalysts that facilitate biochemical reactions in horses by lowering the activation energy required for these processes. They are involved in various physiological functions, including digestion, metabolism, and cellular repair. Common enzymes in equine biology include amylase, lipase, and lactate dehydrogenase, each playing a specific role in the breakdown of nutrients and energy production. The activity and concentration of these enzymes can vary in response to different physiological and pathological conditions, serving as potential indicators in veterinary diagnostics. This page compiles peer-reviewed research studies and scholarly articles that explore the function, regulation, and clinical implications of enzymes in equine health.
Effects of different anticoagulants on determination of erythrocyte glutathione peroxidase. Glutathione peroxidase (GSH-Px) was determined in whole blood from cows, goats and horses using cumenehydroperoxide as substrate. Heparin was found to be the most suitable anticoagulant. The highest activities of GSH-Px were found with high concentrations of heparin in the blood samples (1000 and 1250 IU/ml of blood). Sodium fluoride and especially EDTA and sodium citrate gave lower activities of the enzyme. Storage of the blood samples at room temperature (~20°C) or in a refrigerator (~5°C) for 3 days resulted in significantly lower activities of the enzyme, especially in horse blood. Gluta...
Equine marker genes: Polymorphism for soluble erythrocyte malic enzyme. Polymorphism of equine erythrocyte malic enzyme is detactable on starch gel electrophoresis. The frequency of ME1S was 0.06 in 667 Standardbred and 0.09 in 85 Thoroughbred horses. No genetically determined electrophoretic variation in soluble malate dehydrogenase was detected.
Interaction of horse plasma antithrombin III and alpha 1-proteinase inhibitor with some serine proteinases. Antithrombin III and alpha 1-proteinase inhibitor isolated simultaneously from horse citrated plasma were tested for inhibitory activity against bovine trypsin and chymotrypsin, as well as elastase-like neutral proteinases from horse leucocytes. The stoichiometry of reaction and kinetic parameters (kass, Ko) were estimated and related to the protein pattern obtained after exposure of these proteinases to horse inhibitors as analyzed by polyacrylamide gel electrophoresis (PAGE and PAGE-SDS). As shown by fast reaction rates and low values of dissociation constants the two inhibitors effectively ...
The metabolism of promazine and acetylpromazine in the horse. Promazine hydrochloride and acetylpromazine maleate were administered intravenously at clinical dose levels to horses. In urine from horses given promazine hydrochloride, the parent drug and four metabolites were detected. The two major metabolites, present as conjugates were identified after hydrolysis by beta-glucuronidase/arylsulfatase as 3-hydroxypromazine and 3-hydroxydesmonomethyl-promazine. Conjugated 3-hydroxypromazine has been previously identified as a major metabolite in the horse. Two minor metabolites isolated in this study were primaizine N-oxide and promazine N-oxide sulfoxide. ...
[Some clinical biochemical features of ‘tying up’ in horses (author’s transl)]. The changes in the activities of lactate dehydrogenase (LDH) and creatine phosphokinase (CPK) in de blood of thirty-three horses with 'tying up' were compared. The extent to which the serum enzymes LDH, CPK and glutamic oxaloacetic transaminase (GOT) and the changes in the activities of these enzymes after suitable labour can be used in the diagnosis of 'tying up' and in following the recovery of patients was studied.
Fractionation and partial characterization of alpha-1-protease isoinhibitors of horse. The principal alpha-1-protease inhibitor of horse was fractionated by classical methods and analysed with a modified fibrinogen-agarose gel electrophoretic method of high sensitivity and resolving power. Starting with an electrophoretically homogeneous inhibitor in unfractionated serum, two isoinhibitor bands became apparent after fractionation with (NH4)2SO4 and DEAE-cellulose DE-52 ion-exchange chromatography. The isoinhibitors differed in electrophoretic migration and in the elution pattern from Sephadex G-100 gel filtration, but possessed identical antigenic determinants and enzyme specifi...
Characterization of equine alkaline phosphatase isoenzymes based on their electrophoretic mobility by polyacrylamide gel disc electrophoresis. Alkaline phosphatase isoenzymes of equine tissues, peritoneal fluid, and serum were characterized by their electrophoretic mobilities, using polyacrylamide gel disc electrophoresis. The alkaline phosphatase isoenzymes in liver, kidney, spleen, small intestine, placenta, bone, small colon, and large colon tissue samples were extracted and separated by electrophoresis. The resulting isoenzyme mobilities and spectrophotometric scans were evaluated for their tissue specificity and for their possible use in determining the tissue contribution of alkaline phosphatase to serum and peritoneal fluid. T...
Equine postanesthetic forelimb lameness: intracompartmental muscle pressure changes and biochemical patterns. Intracompartmental muscle pressures were recorded from the right and left forelimbs (extensor carpi radialis, triceps brachii) of healthy horses maintained in left lateral recumbency while under deep halothane anesthesia for 180 to 240 minutes. Cardiac output, blood pressure, blood gases, and acid-base status were monitored throughout the anesthesia, and electrolyte levels (Ca2+, P+, K+, Cl-, Na+) and enzyme activities (aspartate aminotransferase (AST), creatine phosphokinase (CPK), and blood lactate) were monitored for 7 days. Postanesthetic forelimb lameness was produced in 5 of the 6 horses...
Sequence of the low activity equine erythrocyte carbonic anhydrase and delineation of the amino acid substitutions in various polymorphic forms. the sequence of the low activity form of equine erythrocyte carbonic anhydrase has been determined. The most common electrophoretic form, designated D, has been found to have five substitutions. Amino acid exchanges in the electrophoretic variants known as A1, A2, B, and T have been found at six other positions. The data do not permit calculation of the number of polymorphic forms of this enzyme. The equine D isozyme and the analogous human enzyme are quite homologous, 211 of their 260 residues, or 81%, being identical.
Histochemical properties of muscle fibres types and enzyme activities in skeletal muscles of Standardbred trotters of different ages. Fibre characteristics and enzyme activities were determined for the gluteus, semitendinosus, vastus lateralis and triceps brachii muscles of 55 Standardbred trotters of different ages. Four fibre types (I, IIA, IIB, IIC) were demonstrated by histochemical staining of myofibrillar adenosine triphosphatase after preincubation at different pH values. Type II fibres predominated in all the muscles and the type IIA/IIB ratio was higher in horses over 5 years than in younger horses, except in the vastus in which the IIA/IIB ratio did not change with age. The vastus had the highest proportion of type...
Evidence for the existence of two isocolipases in horse pancreas. The N-terminal amino acid sequences of two forms of colipase isolated from horse pancreas have been compared. Four sequence differences were found in the first 51 amino acids. This lead us to conclude that there are two distinct colipases in the horse pancreas.
The complete amino acid sequence of horse muscle acylphosphatase. The amino acid sequence of horse muscle acylphosphatase is given in the present paper. The carboxymethylated enzyme consists of a single polypeptide chain of 98 amino acid residues with an acetyl group blocking the NH2 terminus and a tyrosine at the COOH terminus. The calculated molecular weight of the native protein, a mixed disulfide with glutathione, is 11,365. The carboxymethylated protein was cleaved by cyanogen bromide. The three expected fragments were purified; moreover, an additional fragment, derived from a partial failure of cleavage at methionine-24, was purified and characterized....
Uptake of nucleotides and catecholamines by chromaffin granules from pig and horse adrenal medulla. The uptake of nucleotides and catecholamines into chromaffin granules from adrenals of pigs and horses is similar to that previously seen in bovine chromaffin granules. The rate of [3H]ATP uptake at 2 mM-ATP concentration was 0.42 +/- 0.06 and 0.15 +/- 0.02 nmol/mg protein/min for pig and horse granules, respectively. The apparent Km's were 1.37 mM for pig granules, 0.89 mM for horse granules, and 1.2 mM for ox granules. The sensitivity of the uptake for nucleotides and catecholamine to specific inhibitors was found to be similar in granules from pig and ox, indicating that the same mechanisms...
Plasma concentration of iditol dehydrogenase (sorbitol dehydrogenase) in ponies treated with aflatoxin B1. Twelve clinically normal Shetland ponies were allocated to one of four treatment groups. Aflatoxin B1 was administered at the dosage level of 2 mg/kg of body weight to group A, 1 mg/kg to group B, and 0.5 mg/kg to group C; a placebo was given to group D (controls). Plasma samples were assayed at 4-hour intervals for iditol dehydrogenase (ID) (sorbitol dehydrogenase) concentrations as an indicator of hepatic damage. One of the ponies in group A died 68 hours after dosing; another pony in group A died 76 hours after dosing. All other animals survived the experiment. The means of peak ID values w...
Studies related to the metabolism of anabolic steroids in the horse: the identification of some 16-oxygenated metabolites of 19-nortestosterone. 1. The metabolism of 19-nor[4-14C]testosterone in a thoroughbred horse has been studied and neutral urinary metabolites obtained after enzyme hydrolysis have been investigated by g.l.c.-mass spectrometry. 2. 3-Hydroxyestran-17-one, 17 alpha- and 17 beta-nortestosterone, estrane-3,17-diol (two isomers), 3,16-dihydroxyestran-17-one (two isomers), 3,17-dihydroxyestran-16-one (two isomers) and estrane-3,16,17-triol were identified in the neutral urinary extracts.
Biochemical characterization of equine herpesvirus type 3-induced deoxythymidine kinase purified from lytically infected horse embryo dermal fibroblasts. Infection of horse KyED cells with equine herpesvirus type 3 (EHV-3) resulted in a sevenfold increase in cytosol deoxythymidine kinase (dTK) activity. The EHV-3 dTK was purified from KyED cytosol dTK by affinity chromatography on deoxythymidine-Sepharose and characterized with respect to its electrophoretic mobility, molecular weight, substrate specificity, phosphate donor specificity, and immunological specificity. The purified EHV-3 dTK migrated in polyacrylamide gels with an Rf of 0.30 and sedimented in glycerol gradients with an S value of 5.13, corresponding to a molecular weight of 83,00...
Biochemical effects of succinylcholine chloride in mechanically ventilated horses anesthetized with halothane in oxygen. Succinylcholine chloride administered to horses anesthetized with halothane in oxygen and mechanically ventilated, caused slight but statistically insignificant (P less than 0.01) increases in creatine phosphokinase, lactic dehydrogenase, and aspartate aminotransferase activity. The increases in these enzymes have been explained on the basis of muscle damage resulting from succinylcholine chloride induced muscle fasciculations and by hypoperfusion of tissues due to depression of the cardiovascular system caused by general anesthesia. These changes were not clinically apparent based upon the ab...
The effect of trypsin digestion on the structure and iron-donating properties of transferrins from several species. The effect of trypsin digestion on iron-saturated and iron-free (apo) human, rabbit, bovine, pig and horse tranferrins has been studied. Iron-binding fragments were produced only from iron-saturated pig and bovine transferrins although some cleavage of the polypeptide chain occurred in all cases. The apo-transferrins were generally degraded to a greater extent than the corresponding iron-saturated proteins. The ability of the different transferrins to donate iron to rabbit reticulocytes varied in the order rabbit approximately pig greater than human approximately horse greater than bovine. Try...
Involvement of lysines-72 and -79 in the alkaline isomerization of horse heart ferricytochrome c. Spectrophotometric titrations of five singly modified horse heart ferricytochromes c, specifically (trifluoromethyl)phenylcarbamylated (CF3PhNHCO-) or trifluoroacetylated (CF3CO-) at lysines-13, -72, and -79, were carried out. The CF3PhNHCO-Lys-13, Lys-79, and CF3CO-Lys-79 derivatives all underwent alkaline isomerization with loss of the 695-nm band to low-spin species with an apparent pK of about 8.9, as did the unmodified cytochrome. However, modification of lysine-72 appeared to alter the reaction pathway since the CF3PhNHCO-Lys-72 derivative isomerized to a high-spin form with an apparent ...
Purification by affinity chromatography and characterization of a neutral alpha-glucosidase from horse kidney. A horse kidney neutral alpha-D-glucosidase (alpha-D-glucoside glucohydrolase, EC 3.2.1.20) was purified about 580-fold with a yield of 33% by an affinity chromatography technique using the p-aminophenyl-beta-D-maltoside, a substrate derivative, as ligand. The purified enzyme, homogeneous in polyacrylamide gel electrophoresis, was a glycoprotein with a molecular weight of 280 000 as calculated by gel filtration and its isoelectric focusing points was found to be pH 4.1. The purified enzyme was able to hydrolyze various substrates having (alpha-1,2), (alpha-1,3), (alpha-1,4), and (alpha-1,6) glu...
Methylation of histidine-48 in pancreatic phospholipase A2. Role of histidine and calcium ion in the catalytic mechanism. It is known that His-48 is part of the active center in pancreatic phospholipase. To further elucidate the role of histidine-48 in the active center of pancreatic phospholipase A2, we have modified the enzyme with a number of bromo ketones and methyl benzenesulfonates. Rapid methylation occurred with methyl p-nitrobenzenesulfonate. Methylated phospholipase shows total loss of enzymatic activity whereas binding of substrate and the cofactor Ca2+ remains intact. Amino acid analysis of methylated equine phospholipase showed the loss of the single molecule of histidine and the formation of one mol...
A new method for the isolation of aminoacylase from mammalian kidneys. Aminoacylase (E.C. 3.5.1.14) was isolated from the kidneys of different mammalian species (horse, cattle, rabbit and pig) by extracting the organ with water and subjecting the extract to heat treatment at 70 degrees C for 10 min, then, after having removed denatured proteins by fractionating those remaining in the solution by ammonium sulfate. The enzyme obtained in this way can either be used directly for practical purposes (e.g. preparation of immobilized aminoacylase) or further purified by chromatography. For the further purification of porcine kidney aminoacylase we applied a combination ...
[Bacteriological studies of Haemophilus equigenitalis Taylor 1978, the causative organism of contagious equine metritis 1977 (author’s transl)]. The cultural, biochemical, antigenic and antibiotic susceptibility characteristics of 17 strains of Haemophilus equigenitalis, the causative organism of contagious equine metritis (CEM), were studied. Biochemical characteristics were investigated using both conventional method and the API ZYM system of enzyme detection. The biochemical profile of the H. equigenitalis strains was unique and differed from the other bacterial species studied under the same experimental conditions (H. influenzae and H. parainfluenzae, B. abortus and B. melitensis, P. multocida, A. calcoaceticus). The required X an...