Differential effects of lipids on the osmotic fragility of lysosomes

Summary The effect of a wide range of concentrations of oleic acid, oleyl alcohol and oleic acid methyl ester on lysosomal stability has been studied under both hypotonic and isoosmotic medium conditions. Both oleic acid and oleyl alcohol exhibited a biphasic interaction pattern with lysosomes; stabilizing at low concentrations and labilizing at high concentrations. Lysosome labilization by the ester required an initial lag period.Acknowledgment. We should like to thank Mr.David Ginzburg for the assistance he gave us during the 3 months he spent at our department as a summer student.     

Enzymatic reduction of fatty acids and acyl-CoAs to long chain aldehydes and alcohols

Summary The properties of enzymatic systems involved in the synthesis of long chain aldehydes and alcohols have been reviewed. Fatty acid and acyl-CoA reductases are widely distributed and generate fatty alcohols for ether lipid and was ester synthesis as well as fatty aldehydes for bacterial bioluminescence. Fatty alcohol is generally the major product of fatty acid reduction in crude or membrane systems, although reductases which release fatty aldehydes as products have also been purified. The reduction of fatty acid proceeds through the ATP-dependent formation of acyl intermediates such as acyl-CoA and acyl protein, followed by reduction to aldehyde and alcohol with NAD(P)H. In most cases, both the rate of fatty acid conversion and acyl chain specificity of the reaction are determined at the level of reduction of the intermediate. The reduction of fatty acids represents the major pathway for the control of the synthesis of fatty aldehydes and alcohols. Several other enzymatic reactions involved in lipid degradation also release fatty aldehydes but do not apear to play an important role in long chain alcohol synthesis.     

Enzymatic reduction of fatty acids and acyl-CoAs to long chain aldehydes and alcohols

The properties of enzymatic systems involved in the synthesis of long chain aldehydes and alcohols have been reviewed. Fatty acid and acyl-CoA reductases are widely distributed and generate fatty alcohols for ether lipid and wax ester synthesis as well as fatty aldehydes for bacterial bioluminescence. Fatty alcohol is generally the major product of fatty acid reduction in crude or membrane systems, although reductases which release fatty aldehydes as products have also been purified. The reduction of fatty acid proceeds through the ATP-dependent formation of acyl intermediates such as acyl-CoA and acyl protein, followed by reduction to aldehyde and alcohol with NAD(P)H. In most cases, both the rate of fatty acid conversion and acyl chain specificity of the reaction are determined at the level of reduction of the intermediate. The reduction of fatty acids represents the major pathway for the control of the synthesis of fatty aldehydes and alcohols. Several other enzymatic reactions involved in lipid degradation also release fatty aldehydes but do not appear to play an important role in long chain alcohol synthesis.     

Comparative oxidation of erucic and oleic acids by mitochondria isolated from heart auricle of living man]

Mitochondria were isolated from fragments of heart auricles, that were cut off during surgical intracardiac operations. They were incubated with either [14 14C] erucic acid or [10 14C] oleic acid as a control. In the experimental conditions used, the radioactive products soluble in perchloric acid, that are issued from the beta-oxidation reactions in mitochondria, were formed in much lower amounts from erucic acid than from oleic acid. These results show the very low capacity of human heart mitochondria to use directly erucic acid as a substrate for energy requirements, as has been observed before with other animal species. Activation of fatty acids, the preliminary step of their beta-oxidation, was also observed to be very much lower with erucic acid.     

Cholesterol esterification activities in the intestines and pancreas of the albino rat

Summary Cholesterol esterification activities in intestines and pancreas are much greater with unsaturated fatty acids than with the saturated ones; the maximum activity is with arachidonic acid in intestines and with oleic acid in pancreas. The pancreatic cholesterol esterification activity is higher than the intestinal one.     

Quelques propriétés basiques de la streptomycine

Summary Streptomycin forms an insoluble complex with organic substances of which the isoelectric point is situated in the acid range such as cephalin, oleic, palmitic, and tannic acid.     

The effect of endotoxin on membrane fatty acid composition in BCG-sensitized mice

The effects of endotoxin on mouse liver phospholipid fatty acid composition have been investigated. Administration of endotoxin from Salmonella abortus equi led to a decrease in the polyunsaturated fatty acid content of livers from mice sensitized with Bacille Calmette Guérin (BCG). The content of arachidonic acid fell significantly in both the phosphatidylcholine and phosphatidylinositol fractions whereas in the phosphatidylethanolamine fraction the linoleic acid content was significantly reduced. The polyunsaturated fatty acids were replaced by increased amounts of oleic acid and palmitic acid, leading to a reduction in the polyunsaturated to saturated fatty acid ratio.     

Tierexperimentelle Studien über die Alkoholverbrennung

Summary While in dogs intravenously administered alcohol is eliminated as a straight function of time, it was found that the combustion of alcohol, when given intraperitoneally in rats, depends on the concentration at a given time. Since the dog — like man — belongs to a species with a slow alcohol combustion, while the rat (together with the mouse) has the highest metabolic velocity, it is supposed that the supply of alcohol dehydrogenase might determine not only the metabolic velocity as such but also the rhythm of alcohol combustion: when the ADH supply is low, the combustion capacity limit will be reached at low blood alcohol concentrations and the elimination curve will become linear.     

Occurrence of 3-methoxy octadecanoic acid in yeast lipid

Summary The fatty acid composition of a new strain of the yeastRhodotorula glutinis, grown in molasses, has been studied and found to contain palmitic, stearic, oleic, linoleic and linolenic acids, and small amounts of other constituents. In addition, 3-methoxy octadecanoic acid has been shown to be present in the glycolipid fraction.Partly presented at the National Symposium on Natural Product Chemistry held on February 7–8, 1983 at Bose Institute, Calcutta (India).Acknowledgments. Authors are indebted to Prof. S.C. Bhattacharyya, and Prof. A.K. Barua, Department of Chemistry for providing facilities for this work.     

The effect of endotoxin on membrane fatty acid composition in BCG-sensitized mice

Summary The effects of endotoxin on mouse liver phospholipid fatty acid composition have been investigated. Administration of endotoxin fromSalmonella abortus equi led to a decrease in the polyunsaturated fatty acid content of livers from mice sensitized with Bacille Calmette Guérin (GCG). The content of arachidonic acid fell significantly in both the phosphatidylcholine and phosphatidylinositol fractions whereas in the phosphatidylethanolamine fraction the linoleic acid content was significantly reduced. The polyunsaturated fatty acids were replaced by increased amounts of oleic acid and palmitic acid, leading to a reduction in the polyunsaturated to saturated fatty acid ratio.     

Chemie und Stoffwechsel der Polyenfettsäuren

Summary The chemical structure of the polyenoic fatty acids occuring in organ phosphatides and in fish oils is reviewed. The double bonds of all these polyenoic acids are arranged in divinylmethane pattern. Except some of the C₁₆-polyenoic acids of fish oils, these polyenoic acids belong either to the oleic, linoleic or linolenic acid type and have chain lengths C₁₈, C₂₀, and C₂₂. Polyenoic acids of the oleic acid type are present only in small amounts in phosphatides of mammalian origin. Fish oils are lacking in these but predominantly contain polyenoic acids of the linolenic acid type.Metabolic studies have shown that polyenoic acids of linoleic acid type (e.g. arachidonic acid) originate from linoleic acid and those of the linolenic acid type (e.g. C₂₀-pentaenoic and C₂₂-hexaenoic acid) from linolenic acid-both supplemented exogenous-by extension of the carbon chain by acetate on the side of the carboxylic acid group and introduction of additional double bonds in the divinylmethane pattern directed toward the carboxylic acid group. There is some evidence, however, that a total synthesis of the polyenoic acids of the oleic acid type occurs in the animal body.The transformation of linolenic to C₂₂-hexaenoic acid and some intermediate reactans have been investigated more precisely by means of the tracer method. As far as the biosynthesis of the polyenoic fatty acids is concerned there are no fundamental differences in different vertebrates.

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Nach einem Vortrag auf der gemeinsamen Tagung der deutschen, französischen und schweizerischen Biochemiker in Zürich vom 10.–12. Oktober 1960.     

Comparison in genetically obese and normal rats of the uptake and incorporation of labelled lauric acid, oleic acid, and glycerol by the isolated perfused liver]

Lauric acid, labelled oleic acid and glycerol are perfused in isolated liver of fafa Rats and Wistar Rats previously subjected to fasting. They synthesize TG and PL de novo, though in long time experiments with the normal Rat, the most important method of synthesis is an exchange of AG of the endogenous glycerolipids. However PL are not synthesized with lauric acid. In the livers of fafa Rats the synthesis of TG with oleic acid and glycerol is higher than in livers of Wistar Rats: 16:0 18 : 1 18: 1, 16:0 18: 1 18: 2, 18 : 1 18 :1 18:1, 16 : 0 16 :0 18 : 1 (this TG is not present in liver of Wistar Rat). The hepatic synthesis of PL by the fafa Rat, is less important after 15 min while it is important with Wistar Rats. The synthesized TG with lauric acid (only the TG 12 : 0 12 : 0 12 : 0 with the fafa Rat) are more rapidly oxidized by liver of obese Rat than by liver of normal Rat.     

Sterol carrier protein-2: structure reveals function

The multiple actions of sterol carrier protein-2 (SCP-2) in intracellular lipid circulation and metabolism originate from its gene and protein structure. The SCP-x/pro-SCP-2 gene is a fusion gene with separate initiation sites coding for 15-kDa pro-SCP-2 (no enzyme activity) and 58-kDa SCP-x (a 3-ketoacyl CoA thiolase). Both proteins share identical cDNA and amino acid sequences for 13-kDa SCP-2 at their C-termini. Cellular 13-kDa SCP-2 derives from complete, posttranslational cleavage of the 15-kDa pro-SCP-2 and from partial posttranslational cleavage of 58-kDa SCP-x. Putative physiological functions of SCP-2 have been proposed on the basis of enhancement of intermembrane lipid transfer (e.g., cholesterol, phospholipid) and activation of enzymes involved in fatty acyl CoA transacylation (cholesterol esters, phosphatidic acid) in vitro, in transfected cells, and in genetically manipulated animals. At least four important SCP-2 structural domains have been identified and related to specific functions. First, the 46-kDa N-terminal presequence present in 58-kDa SCP-x is a 3-ketoacyl-CoA thiolase specific for branched-chain acyl CoAs. Second, the N-terminal 20 amino acid presequence in 15-kDa pro-SCP-2 dramatically modulates the secondary and tertiary structure of SCP-2 as well as potentiating its intracellular targeting coded by the C-terminal peroxisomal targeting sequence. Third, the N-terminal 32 amino acids form an amphipathic a-helical region, one face of which represents a membrane-binding domain. Positively charged amino acid residues in one face of the amphipathic helices allow SCP-2 to bind to membrane surfaces containing anionic phospholipids. Fourth, the hydrophobic faces of the N-terminal amphipathic a helices along with beta strands 4, 5, and helix D form a ligand-binding cavity able to accommodate multiple types of lipids (e. g., fatty acids, fatty acyl CoAs, cholesterol, phospholipids, isoprenoids). Two-dimensional 1H-15N heteronuclear single quantum coherence spectra of both apo-SCP-2 and of the 1:1 oleate-SCP-2 complex, obtained at pH 6.7, demonstrated the homogenous formation of holo-SCP-2. While comparison of the apo- and holoprotein amide fingerprints revealed about 60% of the resonances remaining essentially unchanged, 12 assigned amide residues underwent significant chemical-shift changes upon oleic acid binding. These residues were localized in three regions: the juncture of helices A and B, the mid-section of the beta sheet, and the interface formed by the region of beta strands 4, 5, and helix D. Circular dichroism also showed that these chemical-shift changes, upon oleic acid binding, did not alter the secondary structure of SCP-2. The nuclear magnetic resonance chemical shift difference data, along with mapping of the nearby hydrophobic residues, showed the oleic acid-binding site to be comprised of a pocket created by the face of the beta sheet, helices A and B on one end, and residues associated with beta strands 4, 5, and helix D at the other end of the binding cavity. Furthermore, the hydrophobic nature of the previously ill-defined C-terminus suggested that these 20 amino acids may form a 'hydrophobic cap' which closes around the oleic acid upon binding. Thus, understanding the structural domains of the SCP-x/pro-SCP-2 gene and its respective posttranslationally processed proteins has provided new insights into their functions in intracellular targeting and metabolism of lipids.     

Diurnal rhythm of ethanol metabolism in the rat

Male Sprague-Dawley rats injected with 2.0 g/kg of ethanol and analyzed 1 h later at 8 specific times of the day showed diurnal rhythms for alcohol concentrations in the blood, urine, brain and liver tissues. The circadian fluctuation noted for the concentrations of blood and tissue ethanol might indicate a diurnal variation in the enzymatic metabolism of ethanol.     

Diurnal rhythm of ethanol metabolism in the rat

Summary Male Sprague-Dawley rats injected with 2.0 g/kg of ethanol and analyzed 1 h later at 8 specific times of the day showed diurnal rhythms for alcohol concentrations in the blood, urine, brain and liver tissues. The circadian fluctuation noted for the concentrations of blood and tissue ethanol might indicate a diurnal variation in the enzymatic metabolism of ethanol.Supported by a grant from the US National Aeronautics and Space Administration.     

Isolation of indole-3-acetic acid methyl ester,a metabolite of indole-3-acetic acid fromPseudomonas amygdali

From the culture filtrates ofPseudomonas amygdali the methyl ester of indole-3-acetic acid (IAA), a product of indole-3-acetic acid metabolism which has the same auxin activity as the free acid, has been isolated. This is the first report of its occurrence as a microbial metabolite.     

Labile protein-methyl ester: Comparison between chemically and enzymatically synthesized

Summary The rate of hydrolysis of protein-methyl ester, the enzymatic product of S-adenosylmethionine: proteincarboxyl methyltransferase (EC.2.1.1.24) acting on oxidized ribonuclease, was measured at pH 7.1 and 8.6 at 37°C. The half-life of the hydrolysis of the ester is 25 min at pH 7.1, and 4 min at 8.6. The rate of hydrolysis of the enzymatically formed esters at pH 7.0, in 0.1M phosphate buffer, was about 25 times faster than that of esters formed chemically by reaction with methanol in HCl. The lability of the enzymatically synthesized protein-methyl ester suggests that the esterification is specific to sites such that ionization of neighboring amino acid side chains enhances the rate of the hydrolysis.Acknowledgments. This work was supported by Research Grants AM 09603 from the National Institute of Arthritis and Metabolic Diseases, CA 10439 and CA 12226 from the National Cancer Institute, 1-P01-HD-05874 from the National Institute of Child Health and Human Development, National Institutes of Health, GM 20594-03 from National Institute of General Medical Sciences, USA.     

Sintesi dell'acido clorogenico

Summary Quinic acid methyl ester, having the 1.4 and 5 hydroxy-groups suitably blocked, was condensed with the carbonilcaffeic acid chloride.Gradual hydrolysis of the condensation compound gave place to chlorogenic acid.     

Chemical properties of alcohols and their protein binding sites

Alcohols affect a wide array of biological processes including protein folding, neurotransmission and immune responses. It is becoming clear that many of these effects are mediated by direct binding to proteins such as neurotransmitter receptors and signaling molecules. This review summarizes the unique chemical properties of alcohols which contribute to their biological effects. It is concluded that alcohols act mainly as hydrogen bond donors whose binding to the polypeptide chain is stabilized by hydrophobic interactions. The electronegativity of the O atom may also play a role in stabilizing contacts with the protein. Properties of alcohol binding sites have been derived from X-ray crystal structures of alcohol-protein complexes and from mutagenesis studies of ion channels and enzymes that bind alcohols. Common amino acid sequences and structural features are shared among the protein segments that are involved in alcohol binding. The alcohol binding site is thought to consist of a hydrogen bond acceptor in a turn or loop region that is often situated at the N-terminal end of an alpha-helix. The methylene chain of the alcohol molecule appears to be accommodated by a hydrophobic groove formed by two or more structural elements, frequently a turn and an alpha-helix. Binding at these sites may alter the local protein structure or displace bound solvent molecules and perturb the function of key proteins.     

Widespread occurrence of 2-acetylthiazole-4-carboxylic acid in biological material

2-Acetylthiazole-4-carboxylic acid was shown to be widely distributed in all organisms tested, which included members of the eukaryotes, archaebacteria, and eubacteria. This thiazole, which was identified and quantitated as the methyl ester methoxyamine derivative, was found in these organisms at levels of from 27 to 1100 nmol/g dry weight (d.wt) of tissue. On the basis of its widespread occurrence, the levels at which it occurs in these organisms, and its chemical structure, which contains a reactive carbonyl group, it is proposed that this compound is a previously undescribed coenzyme.