ABSTRACT
Studies of aminopropanol dehydrogenase have shown no enzyme activity or very little activity in rat serum and in blood serum of healthy humans. After a single ip dose of CCl4 two enzymatic forms of aminopropanol dehydrogenase, reacting at different pH optima (alkaline and acid), appeared in the blood serum of rats. MFaximum enzyme activity at the alkaline pH appeared about 24 h after CCl4 administration in serum from young rats and after about 12 h in serum from mature rats. Activity values were about 90-150 mumol NAD reduced in 1 min per 1 l serum. Some biochemicl properties of the enzyme reacting at the alkaline pH optimum were studied. Aminopropanol dehydrogenase activity in serum of rats with acute CCl4 poisoning was determined by following the increase in absorbance at 340 nm due to the production of NADH. This enzyme, appearing in the serum after CCl4 poisoning, may be of interest as a potential indicator enzyme for clinical diagnosis.
Subject(s)
Carbon Tetrachloride Poisoning/enzymology , Oxidoreductases/metabolism , Animals , Male , Propanolamines , Rats , SpectrophotometrySubject(s)
Endotoxins/pharmacology , Lactoylglutathione Lyase/metabolism , Liver/enzymology , Lyases/metabolism , Melanoma , Neoplasm Proteins/pharmacology , Skin Neoplasms , Thiolester Hydrolases/metabolism , Animals , Catalase/antagonists & inhibitors , Circadian Rhythm , Cricetinae , Female , In Vitro Techniques , Lactoylglutathione Lyase/antagonists & inhibitors , Male , Mesocricetus , Mice , Neoplasms, ExperimentalABSTRACT
Determinations were made of glyoxalase I and glyoxalase II acitivity in the liver of mice (BDF1 and DBA2 strains) bearing sarcoma 180 and L1210 leukemia in ascites form. A progressive decrease in the both glyoxalase I and glyoxalase II activities to about 40--60% of that of the control groups was observed within the developing period 8--9 days. Test results are interpreted in the light of the postulated role of this enzyme system in cell division and in the tumor development process.
Subject(s)
Lactoylglutathione Lyase/metabolism , Leukemia L1210/enzymology , Liver/enzymology , Lyases/metabolism , Sarcoma 180/enzymology , Thiolester Hydrolases/metabolism , Animals , Cell Division , Glutathione/analogs & derivatives , Lactoylglutathione Lyase/physiology , Leukemia L1210/etiology , Mice , Mice, Inbred Strains , Sarcoma 180/etiology , Thiolester Hydrolases/physiology , Time FactorsABSTRACT
Glyoxalase I bound to Sepharose 4B was used for synthesis of S-lactoyl-glutathione. The bound enzyme does not lose its activity during several months storing and can be used many times for synthesis of S-lactoyl-glutathione. This reaction product can be used as a substrate for glyoxalase II without any further purification.
Subject(s)
Enzymes, Immobilized/metabolism , Glutathione/analogs & derivatives , Lactoylglutathione Lyase/metabolism , Lyases/metabolism , Thiolester Hydrolases/metabolism , Glutathione/chemical synthesis , Lactates/chemical synthesis , Methods , SepharoseABSTRACT
Methylglyoxal in doses over 25 mg/kg injected intravenously in cats and rabbits produces distinct changes in the cardiovascular and respiratory systems, but has no effect on respiration or circulation when injected intraperitoneally even in doses up to 1 g/kg. The effect of MG on blood pressure depends on the species of the animal. The effects of MG are dose-related and dependent on the route of its administration. Biochemical studies showed a significant rise in serum activities of creatine kinase (EC 2-7-3-2), lactate dehydrogenase (EC 1-1-1-27) and aspartate aminotransferase (EC 2-6-1-1-) after intraperitoneal injection of MG in the dose of 200 mg/kg in rabbits and 500 mg/kg in rats. The observed changes probably indicate damage of muscle tissue by MG, presumably as a result of low content of one of the glyoxalases in the muscles of the experimental animals. Elevation of glucose levels by MG was probably an adrenergic effect. These biochemical changes can serve to evaluate toxicity of MG preparations, which exhibit variations probably owing to varying degree of polymerization.
Subject(s)
Aldehydes/poisoning , Pyruvaldehyde/poisoning , Alanine Transaminase/metabolism , Alkaline Phosphatase/metabolism , Amylases/metabolism , Animals , Aspartate Aminotransferases/metabolism , Bicarbonates/blood , Blood Glucose/analysis , Blood Pressure/drug effects , Cats , Creatine Kinase/metabolism , Dose-Response Relationship, Drug , Electrocardiography , Female , Injections, Intraperitoneal , Injections, Intravenous , L-Lactate Dehydrogenase/metabolism , Lactoylglutathione Lyase , Male , Myocardial Contraction/drug effects , Pyruvaldehyde/administration & dosage , Rabbits , Rats , Rats, Inbred Strains , Respiration/drug effects , Species SpecificitySubject(s)
Body Temperature/drug effects , Breast Neoplasms/drug therapy , Etiocholanolone/therapeutic use , Granulocytes/drug effects , Leukocytes/drug effects , Adult , Aged , Breast Neoplasms/metabolism , Etiocholanolone/administration & dosage , Etiocholanolone/metabolism , Etiocholanolone/pharmacology , Female , Fever/chemically induced , Humans , Kinetics , Middle AgedABSTRACT
1. Crude gammadelta-dioxovalerate was synthesized from laevulinate by two different methods and was purified by Sephadex chromatography. Some analytical reactions of the compound are described. 2. gammadelta-Dioxovalerate is a substrate for glyoxalase I and the GSH derivative formed by this enzyme is hydrolysed by glyoxalase II to form d-alpha-hydroxyglutarate. The K(m) of glyoxalase I for gammadelta-dioxovalerate is 1.0x10(-3)m at pH5.8.3. The u.v.-absorption spectrum of thiol ester, synthesized enzymically from gammadelta-dioxovalerate and GSH by glyoxalase I, is almost identical with that for S-lactoylglutathione. Some optical properties of this thiol ester were measured. 4. Attempts to show reversibility of the glyoxalase system reactions with d-alpha-hydroxyglutarate as substrate were unsuccessful. 5. The possible metabolic role of the gammadelta-dioxovalerate reaction is discussed. It is suggested that one of the metabolic functions of the glyoxalase system may be to provide a mechanism for the entry of this compound into the tricarboxylic acid cycle.
Subject(s)
Aldehydes/metabolism , Keto Acids/metabolism , Lyases/metabolism , Thiolester Hydrolases/metabolism , Chemical Phenomena , Chemistry , Chromatography, Paper , Citric Acid Cycle , Glutathione , Glyoxal , Hydrogen-Ion Concentration , Kinetics , Levulinic Acids , Models, Chemical , Optical Rotation , Structure-Activity Relationship , Valerates/isolation & purification , Valerates/metabolismSubject(s)
Antineoplastic Agents/pharmacology , Glyceraldehyde , Neoplasms, Experimental/drug therapy , Animals , Carcinoma, Ehrlich Tumor/drug therapy , Carcinoma, Hepatocellular/drug therapy , Cricetinae , Culture Techniques , Glyceraldehyde/pharmacology , Injections, Intraperitoneal , Leukemia L1210/drug therapy , Liver Neoplasms , Neoplasm Transplantation , Rats , Sarcoma, Yoshida/drug therapyABSTRACT
Glyoxalase I and glyoxalase II (EC. 4.4.1.5 and EC 3.1.2.6) were separated by gel filtration on Sephadex G-75 and G-100. This simple procedure permitted also the partial purification of glyoxalase II. The purification coefficient in a single run from supernatant from beef liver was about 1 : 30 compared with 1 : 15 after the fifth step of purification with classical methods.
