Stabilization of lactate dehydrogenase
Addition of substances such as dimethylsulfoxide, glycerol, and gelatin to aqueous solutions of lactate dehydrogenase maintains enzymes in stable and fully active form when stored at 2 - 6 C.
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Addition of substances such as dimethylsulfoxide, glycerol, and gelatin to aqueous solutions of lactate dehydrogenase maintains enzymes in stable and fully active form when stored at 2 - 6 C.
Male rats, mice, and guinea pigs were starved for 1, 2, or 3 days, and the metabolism of ethylmorphine, p-nitroanisole, and aniline was studied. Results suggest that the oxidative enzyme systems studied are not interdependent, and the pathways studied appear to be species dependent.
With the aid of a simple model membrane system, the properties of cellulose enzymes and of membrane selectivity and pump-like action are considered. The model is based on materials possibly present on a primitive earth, as well as on a membrane able to sort or concentrate these materials. An overview of the model membrane system is presented in terms of how it is constructed, what its properties are, and what to expect in performance characteristics. The model system is shown to be useful for studying the selective and in some cases accelerated transfer of nutrients and metabolites.
Microparticles composed of each of four enzymically synthesized homopolynucleotides and the same lysine-rich proteinoid have been found to influence the condensation of the AMP-anhydrides of each of four amino acids. The conditions of preparation of the particles and other variables of the experiments control the types of reaction. When a period set of conditions was identified empirically, the incorporation favored the amino acids whose present-day codons are related to the homopolynucleotide in the particle.
A quantitative analysis is made of a linear four-step model for photosynthetic molecular oxygen evolution in which each photochemical trapping center or an associated enzyme cycles through five oxidation states. Based on data obtained with isolated chloroplasts, a number of aspects were considered, including the two perturbations which damp the oscillation of the oxygen flash yield in a flash sequence. The kinetics and the mechanism of deactivation was another aspect investigated.
Description of the properties of a carboxypeptidase, termed 'catheptic carboxypeptidase C,' and a dipeptidase, termed 'Ser-Met dipeptidase.' Both were found in highly purified DAP I from either beef spleen or rat liver. Methods are described for the removal or selective inactivation of these contaminating enzymes.
Description of studies of the kinetics of heat inactivation of phosphoglycerate kinase in the soluble fraction from Hydrogenomonas facilis, its extensive purification, and inhibition by adenosine monophosphate (AMP). No evidence was found for an enzyme which catalyzes adenosine-triphosphate-dependent conversion of 3-phosphoglycerate to 1,3-diphosphoglycerate, AMP, and phosphate.
The isozymes considered are designated 'liver type' and 'muscle type' based on the tissue of highest concentration. Electrophoretic analysis shows that the liver type is found in small amounts or is undetectable in all tissues studied except liver. The muscle type is found in skeletal muscles and kidney. Presumptive hybrid enzymes occur at low levels in chicken liver and kidney. The tissue distribution of glyceron-3-P dehydrogenase in several birds capable of sustained flight is different than in chicken.
The response to UV light of several solid proteins and model compounds has been studied in vacuum and at low temperature, using electron paramagnetic resonance techniques. The results indicate that the details of amino acid composition and sequence, and the tertiary structure of a protein are important in determining both the rate of, and the mechanism for, the production of free radicals, and in determining the conditions under which sulfur-type radicals can be produced. The results presented are related to enzyme inactivation and to the UV stability of proteins generally.
Description of an extraction from a Dublin clay loam soil of a colloidal organic matter complex that is urease active and, by X-ray analysis, free of clays. Urease activity in the clay-free precipitates, as in the soil, was not destroyed by the activity of an added proteolytic enzyme, pronase. This is attributed to the circumstance that native soil urease resides in organic colloidal particles with pores large enough for water, urea, ammonia, and carbon dioxide to pass freely, but nevertheless small enough to exclude pronase.
High gravity, cold and starvation elicited similar responses in male Simonson rats. These responses included a decreased rate in body weight gain, increased metabolism of aniline and p-nitroanisole, and no consistent pattern of change in the metabolism of ethylmorphine. Cold and starvation increased the amount of hepatic cytochrome P-450, while hypobaric-hyperoxia caused no change in any of the parameters measured. When 1% acetone was given to the rats in their drinking water, the effects on drug metabolism were similar to those produced by food restriction in that the metabolism of aniline and p-nitroanisole was increased, and the metabolism of ethylmorphine unchanged. The type I binding spectrum of acetone suggests that it is either a substrate, inhibitor, or both for hepatitic oxidative enzymes.
Three independently isolated ultraviolet light sensitive (uvs) mutants of Anacystis nidulans were characterized. Strain uvs-1 showed the highest sensitivity to UV by its greatly reduced photoreactivation capacity following irradiation. Pretreatment with caffeine suppressed the dark-survival curve of strain uvs-1, thus indicating the presence of excision enzymes involved in dark repair. Under 'black' and 'white' illumination, strain uvs-1 shows photorecovery properties comparable with wild-type cultures. Results indicate that strains uvs-1, uvs-35, and uvs-88 are probably genetically distinct UV-sensitive mutants.
Renin is a proteolytic enzyme secreted by the kidney. Techniques for the direct measurement of renin content of human blood are not available at the present time. Two of the best known causes of remediable hypertension can be diagnosed from abnormalities in renin activity and aldosterone production. In renovascular hypertension, renin secretion is increased because of impaired glomerular perfusion. The renin activity assay, when applied in a carefully controlled fashion, is a useful screening test for treatable causes of hypertension.
Commercially available equipment can be modified to provide automated system for assaying drug metabolism by continuous flow-through. System includes steps and devices for mixing drug with enzyme and cofactor in the presence of pure oxygen, dialyzing resulting metabolite against buffer, and determining amount of metabolite by colorimetric method.
A novel method is reported for determining bacterial levels in urine samples, which method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of non-bacterial ATP. After the removal of non-bacterial ATP, the bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay using an enzyme obtained from the firefly.
Statistically significant differences were found between Skylab simulation crews and controls for glycolytic enzymes. The absence of simultaneous controls for the pre- and postchamber analyses leaves the significance of the findings in the crew during these periods indeterminate.
The synthetic production of carbohydrate on a large scale is discussed. Three possible nonagricultural methods of making starch are presented in detail and discussed. The simplest of these, the hydrolysis of cellulose wastes to glucose followed by polymerization to starch, appears a reasonable and economic supplement to agriculture at the present time. The conversion of fossil fuels to starch was found to be not competitive with agriculture at the present time, but tractable enough to allow a reasonable plant design to be made. A reconstruction of the photosynthetic process using isolated enzyme systems proved technically much more difficult than either of the other two processes. Particular difficulties relate to the replacement of expensive energy carrying compounds, separation of similar materials, and processing of large reactant volumes. Problem areas were pinpointed, and technological progress necessary to permit such a system to become practical is described.
The metabolism of winter rye seedlings (Secale cereale, L. cv. Winter) cultured in 99.6% D2O was investigated. Compared with water grown seedlings, the protein content was much lower in the D2O cultured seedlings and the incorporation of H(3)-leucine and H(3)-phenylalanine into medium to high molecular weight proteins was partially blocked. The synthesis of the enzyme peroxidase was also reduced in the D2O plants. Seedlings cultured in D2O incorporate H(3)-thymidine into DNA, but do not take up H(3)-uridine. These results suggest that some of the toxic effects of D2O culture on higher plants can be attributed to a partial block of protein synthesis.