bluuebunny/crossref_metadata_2025 huggingface dataset:
Amino acid metabolism in various fractions of rat-brain homogenates with special reference to the effect of ethanol
Authors: Häkkinen Hertta-Maija , Kulonen E.
Published: August 2015

1. The increase in brain γ-aminobutyrate, glutamate and aspartate and the decrease in brain glutamine that occur when ethanol is administered to rats in vivo could be reproduced by incubating brain homogenates from rats pretreated with ethanol. 2. For the demonstration of the effects of pretreatment with ethanol on the metabolism of γ-aminobutyrate and glutamine, the whole homogenate could be replaced by various supernatant preparations, and even by the soluble protein fraction, which was less active, however. The ‘postmitochondrial’ sediment could likewise mediate the effects of pretreatment with ethanol. 3. When the brain homogenates from control and ethanol-treated rats were allowed to ‘age’ at 2° for more than 7 days, the metabolic difference at incubation could no longer be demonstrated. The capacities of the homogenate from the control rats had changed to resemble those of ethanol-treated rats. 4. Data are given on the effects of the incubation time and of the concentration of homogenate.
Studies of the enzymic mechanism of the metabolism of diethyl 4-nitrophenyl phosphorothionate (parathion) by rat liver microsomes
Authors: Neal R. A.
Published: August 2015

1. The metabolism of parathion by rat liver microsomes is affected by various enzyme inhibitors in a manner quite typical of the ‘mixed-function oxidase’ enzyme systems. 2. With many of these inhibitors (p-chloromercuribenzoate, Cu2+, 8-hydroxyquinoline) the conversion of parathion into diethyl hydrogen phosphorothionate is less inhibited than conversion into diethyl 4-nitrophenyl phosphate (paraoxon). 3. Compounds containing reduced sulphur stimulate the overall metabolism of parathion. However, the conversion of parathion into diethyl hydrogen phosphorothionate is stimulated more than its conversion into paraoxon. 4. The metabolism of parathion to diethyl hydrogen phosphorothionate is also stimulated by EDTA, Ca2+ and Ba2+, but these stimulatory effects are not additive. 5. The electron acceptors FAD, riboflavine, menadione and methylene blue exhibit a concentration-dependent differential inhibition of the metabolism of parathion to diethyl hydrogen phosphorothionate and to paraoxon. 6. The concentration of parathion required for the half-maximal rate of production of diethyl hydrogen phosphorothionate is significantly different from the concentration required for half-maximal rate of production of paraoxon. 7. The results are discussed in terms of either two separate enzyme systems metabolizing parathion to diethyl hydrogen phosphorothionate and to paraoxon or two different binding sites for parathion, which share a common electron-transport pathway.
Tracer studies on the biosynthesis of amino acids from lactate by Peptostreptococcus elsdenii
Authors: Somerville H. J. , Peel J. L.
Published: August 2015

Peptostreptococcus elsdenii, a strict anaerobe from the rumen, was grown on a medium containing yeast extract and [1−14C]- or [2−14C]-lactate. Radioisotope from lactate was found in all cell fractions, but mainly in the protein. The label in the protein fraction was largely confined to a few amino acids: alanine, serine, aspartic acid, glutamic acid and diaminopimelic acid. The alanine, serine, aspartic acid and glutamic acid were separated, purified and degraded to establish the distribution of 14C from lactate within the amino acid molecules. The labelling patterns in alanine and serine suggested their formation from lactate without cleavage of the carbon chain. The pattern in aspartic acid suggested formation by condensation of a C3 unit derived directly from lactate with a C1 unit, probably carbon dioxide. The distribution in glutamic acid was consistent with two possible pathways of formation: (a) by the reactions of the tricarboxylic acid cycle leading from oxaloacetate to 2-oxoglutarate, followed by transamination; (b) by a pathway involving the reaction sequence 2 acetyl-CoA→crotonyl-CoA→glutaconate→glutamate.
The hydrolysis of algal galactans by enzymes from a Cytophaga species
Authors: Turvey J R , Christison J.
Published: August 2015

1. Two bacteria were isolated from sea water by the enrichment culture technique, both of which could utilize the galactan sulphate, porphyran, as sole source of carbon. 2. From the cells of one bacterium, classified as a Cytophaga sp., hydrolytic enzymes were isolated. 3. Partial purification of the enzymes is described and some of the properties of the principal enzymes have been studied. 4. The action of the enzymes on several galactan sulphates of red algae suggests that an agarase is present in the mixture.
Studies of intermediary metabolism in germinating pea cotyledons. The pathway of ethanol metabolism and the role of the tricarboxylic acid cycle
Authors: Cameron D. S. , Cossins E. A.
Published: August 2015

1. The pathway of ethanol metabolism in cotyledons of 3-day-old pea seedlings has been examined by incubating tissue slices with [1−14C]ethanol and [2−14C]ethanol for periods up to 1hr. 2. Ethanol was rapidly incorporated into citrate and glutamate but relatively small amounts of 14C were present in the evolved carbon dioxide even after 1hr. of ethanol metabolism. 3. Similar data were obtained from experiments in which [1,2−14C2]acetaldehyde and [14C]acetate were supplied. 4. The results are interpreted as indicating that ethanol is metabolized essentially via the reactions of the tricarboxylic acid cycle with a substantial drain of α-oxoglutarate to support the biosynthesis of glutamate. 5. It is concluded that oxaloacetate, required for the incorporation of ethanol into citrate, arises mainly from the transamination of aspartate and the fixation of carbon dioxide.
The biosynthesis of 5-hydroxytryptamine in brain
Authors: Grahame-Smith D. G.
Published: August 2015

1. Studies on the anatomical distribution of tryptophan 5-hydroxylase in dog brain show that the activity of this enzyme parallels the activity of 5-hydroxytryptophan decarboxylase and the concentration of 5-hydroxytryptamine in various areas of the brain. 2. Subcellular fractionation of homogenates of rabbit hind-brain has shown that at least 40% of the tryptophan 5-hydroxylase activity is associated with the crude mitochondrial fraction; equilibrium centrifugation in a discontinuous sucrose gradient suggests that this particulate tryptophan 5-hydroxylase is localized in the synaptosomes (nerve-ending particles). 3. Attempts to increase the activity of tryptophan 5-hydroxylase in iso-osmotic homogenates of brain by adding 5,6,7,8-tetrahydro-6,7-dimethylpteridine failed. 4. Procedures designed to rupture synaptosomes caused a fall in the activity of tryptophan 5-hydroxylase, which fall was reversed by adding 5,6,7,8-tetrahydro-6,7-dimethylpteridine. This suggested either that the intact synaptosome, in which the tryptophan 5-hydroxylase is localized, contained an optimum amount of the pteridine, or that the pteridine could not pass across the nerve-cell membrane. 5. Partial purification of tryptophan 5-hydroxylase from brain preparations by ammonium sulphate precipitation revealed that the enzyme was completely dependent on 5,6,7,8-tetrahydro-6,7-dimethylpteridine for activity.
3-Methylhistidine in actin and other muscle proteins
Authors: Johnson P. , Harris C I , Perry S V
Published: August 2015

1. By the use of the extended elution system for basic amino acid analysis, 3-methylhistidine has been detected in hydrolysates of actin isolated from mammalian, fish and bird skeletal muscle. 2. Evidence is presented to indicate that 3-methylhistidine forms part of the primary structure and that in rabbit actin this residue is restricted to one peptide fraction obtained from the tryptic digest. 3. Rabbit skeletal-muscle actin has a 3-methylhistidine:histidine ratio 1:7·6, indicating a minimum molecular weight of 47600. 4. Adult rabbit myosin contains approximately 2 3-methylhistidine residues/mol. These residues are localized in the heavy meromyosin part of the molecule, and are restricted to the major component obtained after succinylation.
Stability of unimolecular films of 32P-labelled lecithin
Authors: Hauser H. , Dawson R. M. C.
Published: August 2015

1. The stability of monolayers of a highly unsaturated yeast lecithin labelled with 32P has been investigated by a surface radioactivity technique. 2. Lecithin films on distilled water at all surface pressures between 6 and 48dynes/cm. were completely stable on rapid perfusion of the subphase and on addition of ionic amphipathic substances to the film. 3. Ultrasonically treated lecithin added to the subphase caused a slow loss of surface radioactivity but little pressure change. 4. The addition of proteins to the subphase caused negligible changes in the film even when conditions were favourable for electrostatic heterocoagulation and penetration. 5. Lecithin films were not hydrolysed by a strongly acid subphase at room temperature. The very low rate of hydrolysis produced by alkali was proportional to the subphase OH−ion concentration: the apparent activation energy and temperature coefficient (Q10) of the reaction were 14250 cal. and 2·37 respectively. 6. Alkaline hydrolysis of lecithin monolayers was markedly stimulated by adding methanol (10–20%, v/v) to the subphase. The addition of ionic amphipaths to the monolayer had the expected type of effect on the hydrolysis rate, but its magnitude was far less than that suggested by an application of the Poisson–Boltzmann equation for ion distribution at a charged interface (Davies & Rideal, 1963).
The purification and properties of isocitrate lyase from Chlorella
Authors: John P. C. L. , Syrett P. J.
Published: August 2015

1. Isocitrate lyase (threo-ds-isocitrate glyoxylate-lyase, EC 4.1.3.1) has been purified from acetate-adapted cells of Chlorella pyrenoidosa. 2. The final preparation was homogeneous by the criteria of sedimentation, diffusion and polyacrylamide-gel electrophoresis. 3. The sedimentation coefficient (S20,w) was 9·04×10−13sec. and the diffusion coefficient (D20,w) 4·62×10−7cm.2/sec.; from these values the molecular weight of the enzyme was calculated to be 170000 and its Stokes radius to be 4·63×10−7cm. 4. The elution of the enzyme from Sephadex G-100 was studied and estimates of molecular weight and Stokes radius were obtained from the elution data. 5. The turnover number of the enzyme was 5950moles of glyoxylate formed/min./mole of enzyme at 30°. 6. With threo-ds(+)-isocitrate as substrate, the Km of the enzyme was 0·023mm.
The biosynthesis of cytochrome c. Sequence of incorporation in vivo of [14C]lysine into cytochrome c and total proteins of rat-liver subcellular fractions
Authors: González-Cadavid N. F. , Campbell P. N.
Published: August 2015

1. In order to determine the initial intracellular site of synthesis of cytochrome c in the liver cell, groups of rats were injected with [14C]lysine and killed 7·5, 15, 30 and 60min. later. The livers were homogenized in 0·3m-sucrose and subcellular fractions obtained. The mitochondrial fraction was further subfractionated. Pure cytochrome c was isolated from extracts of each fraction, obtained first with water at pH4·0 and then with 0·15m-sodium chloride. 2. A comparison of the kinetics of incorporation of [14C]lysine into total protein for each particulate fraction showed the usual two different kinds of kinetics. Incorporation into all the mitochondrial subfractions and the nuclear fraction rose gradually to a plateau value at about 20min., in contrast with that into the two microsomal fractions which rose rapidly to a peak value about seven times that for the mitochondrial fractions. The kinetics for the incorporation into mitochondrial cytochrome c showed a plateau value at 30min. about three times that for the total mitochondrial protein. There was no difference in the specific radioactivity of the mitochondrial cytochrome c extracted with water or 0·15m-sodium chloride or between the different mitochondrial subfractions. In contrast, the cytochrome c isolated from water extracts of the microsomal fractions had a lower specific radioactivity than that obtained from the 0·15m-sodium chloride extract. The specific radioactivity of the latter showed a rapid rise to a peak value about four times that for the mitochondrial cytochrome c, and the shape of the curve was similar to that for the total protein of the microsomal fraction. The results suggest that cytochrome c is synthesized in toto by the morphological components of the microsomal fraction. It seems first to be bound tightly to a microsomal particle, passing then to a looser microsomal binding and being finally transferred to the mitochondria. The newly synthesized cytochrome c in the mitochondrion could not be differentiated from the old by its degree of extractability at pH 4·0.
Esters of serine and threonine in hydrolysates of histones and protamines, and attendant errors in amino acid analyses of proteins
Authors: Murray K. , Milstein C.
Published: August 2015

1. Partial acid hydrolysates of histones from various origins and of protamine were analysed by a two-dimensional ionophoretic procedure to reveal strongly acidic ninhydrin-positive components. 2. Histone fractions prepared by extraction with sulphuric acid gave rise to spots identified as serine O-sulphate and threonine O-sulphate. These two compounds, which were not found in hydrolysates of corresponding fractions prepared by extraction with hydrochloric acid, were artifacts. 3. Hydrolysis of proteins in the presence of traces of sulphate can lead to the formation of the O-sulphates of serine and threonine. This can cause errors, which may sometimes be serious, in amino acid analyses of proteins. 4. O-Phosphoserine was obtained in small amounts from some histone fractions and from protamine, but was undetectable in other histone fractions, notably those of lower lysine content.
Phosphorylation of choline and ethanolamine in Ehrlich ascites-carcinoma cells
Authors: Sung Cheng-Po , Johnstone R. M.
Published: August 2015

1. Ehrlich ascites-cell extracts convert choline and ethanolamine approximately equally well into their respective phosphoryl derivatives. 2. Choline is a potent inhibitor of ethanolamine phosphorylation, but ethanolamine has little effect on choline phosphorylation. 3. 2,3-Dimercaptopropanol, cysteine and Ca2+ inhibit ethanolamine phosphorylation, but have no detectable effect on choline phosphorylation. 4. Choline-phosphorylating activity in Ehrlich ascites-cell extracts is more stable during storage than ethanolamine-phosphorylating activity. 5. Choline phosphorylation is stimulated in the presence of benzoylcholine, succinylcholine, butyrylcholine and propionylcholine, whereas ethanolamine phosphorylation is inhibited. This relationship is reciprocal: the compounds causing the greatest stimulation of choline phosphorylation bring about the greatest inhibition of ethanolamine phosphorylation.
The determination of lactate dehydrogenase isoenzymes in normal human muscle and other tissues
Authors: Emery A. E. H.
Published: August 2015

1. A technique has been developed, based on preferential inhibition by urea, for determining the amounts and proportions of the M and H sub-units of lactate dehydrogenase (referred to as LDH-M and LDH-H respectively) in human tissues, including muscle. 2. There was good agreement between the results obtained with urea inhibition and those obtained with starch-gel electrophoresis. 3. With increasing age there was a significant decrease in the total amount of lactate dehydrogenase and the amount of LDH-M in skeletal muscle. This could not be accounted for by the replacement of functioning muscle tissue by fibrous connective tissue. 4. The proportion of LDH-M was less in certain muscles (e.g. soleus and extra-ocular) than in other muscles (e.g. gastrocnemius and rectus abdominis). 5. The proportions of LDH-M and LDH-H did not differ significantly in different superficial limb muscles and were not significantly affected by either age or sex. 6. Specimens of muscle from 86 different individuals (all Europeans) have been subjected to electrophoresis, but no variants of lactate dehydrogenase isoenzymes have been found.
A simple method for the preparation of 5-methyltetrahydropteroylglutamic acid
Authors: Chanarin I. , Perry Janet
Published: August 2015

Acidification of 5-formyltetrahydropteroylglutamic acid, followed by reduction at pH7·0, provided a simple method of preparing 5-methyltetrahydropteroylglutamic acid.
Binding of nicotinamide–adenine dinucleotides to diphtheria toxin
Authors: Montanaro L. , Sperti Simonetta
Published: August 2015

1. Changes in protein fluorescence have been utilized in determining the stoicheiometry and dissociation constants of the complexes of diphtheria toxin with NADH2, NAD, NADPH2 and NADP. 2. The binding stoicheiometry is 2moles of NADH2 and 1mole of NADPH2/mole of diphtheria toxin. The binding sites for NADH2 appear to be equivalent and independent. 3. The toxin shows a higher affinity for the reduced than for the oxidized forms of the nucleotides. 4. Dissociation constants at 0·01I, pH7 and 25° are 0·7×10−6m for NADH2 and 0·45×10−6m for NADPH2. Dissociation constants increase with increasing ionic strength, indicating that the binding is mainly electrostatic. 5. Bound NADH2 and NADPH2 may be activated to fluoresce by the transfer of energy from the excited aromatic amino acids of the toxin. Activation and emission spectra of bound and free nucleotides are compared. 6. Since NAD and NADH2 are cofactors specifically required for the inhibition of protein synthesis by diphtheria toxin, the possible role of toxin–nucleotide complexes is discussed in this regard.
The extraction of cell walls of Pseudomonas aeruginosa with aqueous phenol. The insoluble residue and material from the aqueous layers
Authors: Clarke K. , Gray G. W. , Reaveley D. A.
Published: August 2015

1. The insoluble residue and material present in the aqueous layers resulting from treatment of cell walls of Pseudomonas aeruginosa with aqueous phenol were examined. 2. The products (fractions AqI and AqII) isolated from the aqueous layers from the first and second extractions respectively account for approx. 25% and 12% of the cell wall and consist of both lipopolysaccharide and muropeptide. 3. The lipid part of the lipopolysaccharide is qualitatively similar to the corresponding material (lipid A) from other Gram-negative organisms, as is the polysaccharide part. 4. The insoluble residue (fraction R) contains sacculi, which also occur in fraction AqII. On hydrolysis, the sacculi yield glucosamine, muramic acid, alanine, glutamic acid and 2,6-diaminopimelic acid, together with small amounts of lysine, and they are therefore similar to the murein sacculi of other Gram-negative organisms. Fraction R also contains substantial amounts of protein, which differs from that obtained from the phenol layer. 5. The possible association or aggregation of lipopolysaccharide, murein and murein sacculi is discussed.
Citrate and the conversion of carbohydrate into fat. The regulation of fatty acid synthesis by rat liver extracts
Authors: Fang Marie , Lowenstein John M.
Published: August 2015

1. The rate of fatty acid synthesis by particle-free extracts prepared from rat liver is increased greatly if the enzyme system is first activated with citrate. 2. The extent of the activation depends on the citrate concentration and on the time of activation in an interdependent manner. 3. Citrate activation is strongly dependent on temperature. 4. Tricarballylate can replace citrate as an activator, but its presence in the assay inhibits fatty acid synthesis. 5. Mg2+ ions can replace citrate in the activation but not in the complete reaction system. 6. ATP prevents the activating effect of citrate and Mg2+ ions. 7. The rate of fatty acid synthesis is increased by palmitoyl-dl-carnitine. This type of activation, additional to that caused by citrate, is rapid and does not depend on prior incubation. 8. Inhibition of fatty acid synthesis by palmitoyl-CoA can be prevented by palmitoyl-dl-carnitine or by increasing the concentration of protein.
Resting and stimulated respiration in vitro in the central nervous system. Regional distribution and relation to cell density
Authors: Ridge J. W.
Published: August 2015

1. The respiratory rates in vitro of ten structures of the rabbit brain were measured in the presence of a normal (5mm) and an elevated (50mm) concentration of K+ ions. The results were expressed on a dry-weight basis and in terms of cell density. 2. On a dry-weight basis, with a normal concentration of K+, there was a steady decrease in respiratory rate on passing from the cerebral cortex through successively lower centres to the brain-stem, i.e. respiration was negatively correlated with phylogenetic age. 3. When the resting respiratory rates were expressed in terms of cell density there was no correlation with phylogenetic age. The neuron-containing structures of the cerebrum and brain-stem had identical respiratory rates with the exception of the cerebral cortex which had a higher rate. 4. The K+-stimulated respiratory rates/cell also showed a negative correlation with phylogenetic age; the higher centres responded with a greater percentage increase than the lower. 5. The results from the cerebellar cortex were anomalous. 6. There were variations of respiratory rate within the diencephalon. The thalamus had a relatively high respiratory rate, the anterior and ventral regions relatively low and the hypothalamus intermediate. 7. The oxygen consumption of white matter was not increased by a high external concentration of K+.
The aminoethylphosphonate-containing lipids of rumen protozoa
Authors: Dawson R. M. C. , Kemp P
Published: August 2015

1. A method is presented for identifying and estimating the aminoethylphosphonate (ciliatine)-containing phospholipids in a complex mixture. 2. Evidence was obtained that the phospholipids of a pure culture of Entodinium caudatum and a mixed rumen protozoa sample contain diglyceride ciliatine, and a plasmalogen ciliatine was detected in the latter. 3. A ninhydrin-positive sphingolipid was isolated from rumen protozoa. Although chromatographically homogeneous on silica gel it contains two components, which were provisionally identified as ceramide ciliatine and ceramide phosphorylethanolamine. 4. A detailed phospholipid analysis of E. caudatum and rumen protozoa is presented. They contain no phosphatidylserine or cardiolipin, but an unidentified phosphoglyceride containing a zwitterionic amino acid is present.
Carbohydrate metabolism of the perfused rat liver
Authors: Ross B. D. , Hems R. , Freedland R. A. , Krebs H. A.
Published: August 2015

1. The rates of gluconeogenesis from most substrates tested in the perfused livers of well-fed rats were about half of those obtained in the livers of starved rats. There was no difference for glycerol. 2. A diet low in carbohydrate increased the rates of gluconeogenesis from some substrates but not from all. In general the effects of a low-carbohydrate diet on rat liver are less marked than those on rat kidney cortex. 3. Glycogen was deposited in the livers of starved rats when the perfusion medium contained about 10mm-glucose. The shedding of glucose from the glycogen stores by the well-fed liver was greatly diminished by 10mm-glucose and stopped by 13·3mm-glucose. Livers of well-fed rats that were depleted of their glycogen stores by treatment with phlorrhizin and glucagon synthesized glycogen from glucose. 4. When two gluconeogenic substrates were added to the perfusion medium additive effects occurred only when glycerol was one of the substrates. Lactate and glycerol gave more than additive effects owing to an increased rate of glucose formation from glycerol. 5. Pyruvate also accelerated the conversion of glycerol into glucose, and the accelerating effect of lactate can be attributed to a rapid formation of pyruvate from lactate. 6. Butyrate and oleate at 2mm, which alone are not gluconeogenic, increased the rate of gluconeogenesis from lactate. 7. The acceleration of gluconeogenesis from lactate by glucagon was also found when gluconeogenesis from lactate was stimulated by butyrate and oleate. This finding is not compatible with the view that the primary action of glucagon in promoting gluconeogenesis is an acceleration of lipolysis. 8. The rate of gluconeogenesis from pyruvate at 10mm was only 70% of that at 5mm. This ‘inhibition’ was abolished by oleate or glucagon.
The cellular distribution of some rat-kidney glycosidases
Authors: Price R. G. , Dance N.
Published: August 2015

1. Free and total activities of β-glucosidase, β-galactosidase, N-acetyl-β-glucosaminidase and β-glucuronidase have been determined fluorimetrically in five subcellular fractions of rat kidney. 2. The β-glucosidase activity appeared in the soluble fraction, β-glucuronidase had the distribution pattern of a lysosomal enzyme, and both β-galactosidase and N-acetyl-β-glucosaminidase had bimodal distributions. 3. Two types of β-galactosidase activity were found: a sedimentable type, having optimum pH3·7, mol.wt. about 80000 and slow electrophoretic mobility at pH7·0 in starch gel; and a soluble type of much faster mobility, having optimum pH5·5–6·5 and mol.wt. about 40000. 4. Evidence is presented that the β-glucosidase and the soluble type of β-galactosidase are the same enzyme. 5. Most of the N-acetyl-β-glucosaminidase activity was in the lysosome-rich fractions, but a significant proportion occurred in the microsomal fraction in a non-latent form. 6. The use of β-galactosidase and N-acetyl-β-glucosaminidase as lysosomal marker enzymes is complicated by the possible presence of multiple forms, but this limitation does not apply to β-glucuronidase in the rat kidney.
The intracellular ratio of cysteine and cystine in various tissues
Authors: Crawhall J. C. , Segal S.
Published: August 2015

1. The cysteine–cystine ratio was measured in rat kidney cortex, diaphragm, jejunum, liver and brain. 2. This ratio was determined by incubating these tissues in buffer containing [35S]cystine and then homogenizing the tissue in a buffered solution of N-ethylmaleimide. The products of this reaction were separated by high-voltage electrophoresis and the radioactivity in the cystine and 2-(l-2′-amino-2′-carboxyethylthio)-N-ethylsuccinimide regions was determined. 3. In these tissues cyst(e)ine was mainly present in the reduced form. 4. After incubation of [35S]cystine with rat jejunal segments it was found that 36% of the cystine in the medium has been reduced. 5. Anaerobiosis, Na+-free media, glucose and high concentrations of cystine and lysine were found not to affect significantly the cysteine–cystine ratio in rat kidney-cortex slices.
Chromophores of allophycocyanin and R-phycocyanin
Authors: Chapman D. J. , Cole W. J. , Siegelman H. W.
Published: August 2015

The biliprotein allophycocyanin was purified from Phormidium luridum, Anabaena variabilis and Plectonema boryanum. R-phycocyanin was purified from Rhodymenia palmata. The chromophores were cleaved from the denatured protein by methanol hydrolysis. They were purified and crystallized as the dimethyl esters. Chromatographic and absorption-spectral (visible–ultraviolet and infrared) comparisons with reference material have established phycocyanobilin as the chromophore of allophycocyanin. Phycocyanobilin and phycoerythrobilin were shown to be the chromophores of R-phycocyanin.
Metabolism of the reserve polysaccharide of Streptococcus mitis. Properties of α-(1→6)-glucosidase, its separation from transglucosylase, and the action of the two enzymes on branched oligosaccharides
Authors: Walker Gwen J. , Builder Janet E.
Published: August 2015

1. An α-(1→6)-glucosidase has been separated from cell extracts of Streptococcus mitis. The enzyme was freed from transglucosylase by adsorption of the latter on retrograded amylose. 2. The enzyme was detected in five of the six strains of S. mitis that were studied; α-(1→6)-glucosidase was not found in strain RB1633, a strain that did not store polysaccharide. 3. The glucosidase could act on compounds in which α-glucose is joined through an α-(1→6)-bond to either a maltosaccharide or an isomaltosaccharide. 62-α-Glucosylmaltose (panose) and 63-α-glucosylmaltotriose were hydrolysed more rapidly and isomaltodextrins more slowly than isomaltose. 4. Transferring activity towards isomaltose and panose was appreciable when the concentration of substrate was 2% or higher. 5. The enzyme had no action on α-(1→4)-glucosidic linkages. 6-α-Maltodextrinylglucoses were hydrolysed only after transglucosylase action had attenuated them to isomaltose.
The participation of phospholipids in the interaction of leucocidin and the cell membrane of the polymorphonuclear leucocyte
Authors: Woodin A. M. , Wieneke Antonnette A.
Published: August 2015

1. The interaction of the two components of leucocidin with various lipids has been studied by sedimentation, flotation, light-scattering and changes in the biological activity of leucocidin. 2. Phosphatidylserine, phosphatidylcholine, diphosphoinositide, triphosphoinositide and phosphatidic acid, but not phosphatidylethanolamine, lysophosphatidylcholine, cerebrosides, gangliosides or tristearin, induce aggregation of the F component of leucocidin. 3. The S component of leucocidin does not interact directly with these phospholipids, but interacts with the F component of leucocidin after its modification by lipids. 4. The increased sedimentation or light-scattering induced by low phospholipid concentrations is reversed at higher phospholipid concentrations. 6. The aggregates formed by phospholipids and leucocidin are due, not to adsorption of leucocidin alone, but also to the formation of leucocidin polymers. 7. It is concluded that the aggregation is due to the interaction of the F component with the fatty acid side chains in the lipid micelle. 8. The S component of leucocidin is inactivated by triphosphoinositide at physiological ionic strength; the F component of leucocidin is inactivated at low ionic strength by triphosphoinositide and remains inactive when the ionic strength is increased. 9. It is suggested that in the leucocyte cell membrane the S component of leucocidin interacts with the polar hydrophilic groups of triphosphoinositide and that the F component of leucocidin interacts with the hydrophobic parts of triphosphoinositide.
The polyphosphoinositide content of the leucocyte, erythrocyte and macrophage
Authors: Wieneke Antonnette A. , Woodin A. M.
Published: August 2015

1. The polyphosphoinositide content of macrophages and the cell membranes of leucocytes and erythrocytes was determined by an extension of the ‘acid-hydrolysis’ procedure of Dawson & Eichberg (1965). The estimation was controlled by adding a little highly radioactive polyphosphoinositide to the tissue extracts before fractionation. Several standard methods for determining polyphosphoinositides gave low recoveries when applied to leucocytes, and it is suggested that these cells contain materials that form complexes with the polyphosphoinositides and interfere with the assay. 2. The method for the preparation of leucocyte cell surface membranes has been modified.
The effect of diet on carbon tetrachloride metabolism
Authors: Seawright A. A. , McLean A. E. M.
Published: August 2015

1. Blood and liver concentrations of carbon tetrachloride were measured, at intervals after an oral dose, in rats given stock and protein-free diets. The values did not correlate with the resistance to poisoning found in the rats on protein-free diets. 2. The metabolism of carbon tetrachloride to carbon dioxide in vivo and in liver microsomal preparations was depressed in animals given protein-free diets. 3. Rats given a single dose of DDT [1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane] were highly sensitive to carbon tetrachloride poisoning. The livers of such animals had an increased microsomal protein content and greatly increased microsomal activity in the demethylation of Pyramidon (aminopyrine) and in the conversion of 14CCl4 into 14CO2. 4. The incorporation of [14C]leucine into protein by liver slices was depressed by carbon tetrachloride. This effect was decreased by addition of SKF525A (2-diethylaminoethyl 2,2-diphenyl-2-propylacetate) and in slices from rats given protein-free diets. It is suggested that the toxicity of carbon tetrachloride is closely linked to its metabolism.
Respiration-driven proton translocation in rat liver mitochondria
Authors: Mitchell Peter , Moyle Jennifer
Published: August 2015

1. Pulses of acidity of the outer aqueous phase of rat liver mitochondrial suspensions induced by pulses of respiration are due to the translocation of H+ (or OH−) ions across the osmotic barrier (M phase) of the cristae membrane and cannot be attributed to the formation (with acid production) of a chemical intermediate that subsequently decomposes. 2. The effective quantity of protons translocated per bivalent reducing equivalent passing through the succinate-oxidizing and β-hydroxybutyrate-oxidizing spans of the respiratory chain are very close to 4 and 6 respectively. These quotients are constant between pH5·5 and 8·5 and are independent of changes in the ionic composition of the mitochondrial suspension medium provided that the conditions permit the accurate experimental measurement of the proton translocation. 3. Apparent changes in the →H+/O quotients may be induced by conditions preventing the occurrence of the usual backlash; these apparent changes of →H+/O are attributable to a very fast electrically driven component of the decay of the acid pulses that is not included in the experimental extrapolations. 4. Apparent changes in the →H+/O quotients may also be induced by the presence of anions, such as succinate, malonate and phosphate, or by cations such as Na+. These apparent changes of →H+/O are due to an increase in the rate of the pH-driven decay of the acid pulses. 5. The uncoupling agents, 2,4-dinitrophenol, carbonyl cyanide p-trifluoromethoxyphenylhydrazone and gramicidin increase the effective proton conductance of the M phase and thus increase the rate of decay of the respiration-driven acid pulses, but do not change the initial →H+/O quotients. The increase in effective proton conductance of the M phase caused by these uncouplers accounts quantitatively for their uncoupling action; and the fact that the initial →H+/O quotients are unchanged shows that uncoupler-sensitive chemical intermediates do not exist between the respiratory-chain system and the effective proton-translocating mechanism. 6. Stoicheiometric acid–base changes associated with the activity of the regions of the respiratory chain on the oxygen side of the rotenone- and antimycin A-sensitive sites gives experimental support for a suggested configuration of loop 3.
The conversion of cholest-7-en-3β-ol into cholesterol. General comments on the mechanism of the introduction of double bonds in enzymic reactions
Authors: Dewhurst S. Marsh , Akhtar M
Published: August 2015

Convenient syntheses of 6β-tritiated Δ7-cholestenol and 3α-tritiated Δ7-cholestene-3β,5α-diol are described. It was shown that the conversion of 6β-tritiated Δ7-cholestenol into cholesterol is accompanied by the complete retention of label. It was unambiguously established that the overall reaction leading to the introduction of the double bond in the 5,6-position in cholesterol occurs via a cis-elimination involving the 5α- and 6α-hydrogen atoms and that during this process the 6β-hydrogen atom remains completely undisturbed. Metabolic studies with 3α-tritiated Δ7-cholestene-3β,5α-diol revealed that under anaerobic conditions the compound is not converted into cholesterol. This observation, coupled with the previous work of Slaytor & Bloch (1965), is interpreted to exclude a hydroxylation–dehydration mechanism for the origin of the 5,6-double bond in cholesterol. It was also shown that under aerobic conditions 3α-tritiated Δ7-cholestene-3β,5α-diol is efficiently converted into cholesterol and that this conversion occurs through the intermediacy of 7-dehydrocholesterol. Cumulative experimental evidence presented in this paper and elsewhere is used to suggest that the 5,6-double bond in cholesterol originates through an oxygen-dependent dehydrogenation process and a hypothetical mechanism for this and related reactions is outlined.
Formation of protoporphyrin from haemoglobin in vitro
Authors: Veech R. L. , Rogeness G. A. , Weil-Malherbe H.
Published: August 2015

1. The formation of protoporphyrin from red blood cells or purified haemoglobin in aqueous perchloric acid media without the prior isolation of haemin is described. The reaction is carried out in the absence of oxygen and in red light. Even traces of oxygen inhibit the reaction by oxidative destruction of protoporphyrin and by the oxidation of haem to haematin. 2. Perchloric acid releases iron and protoporphyrin from haemoglobin at similar rates, but the amount of protoporphyrin in the filtrate varies with the solubility of protoporphyrin in the concentration of perchloric acid used. The yield of protoporphyrin may reach 50–60%. Less than 5μg. of haemoglobin/ml. can be detected by measuring the fluorescence of the porphyrin released. 3. A porphyrin other than protoporphyrin is obtained in small amounts. Its possible identity is discussed. 4. If sodium sulphite is present as a reducing agent the exclusion of oxygen is not required, but the porphyrin formed is more polar and more soluble in water than protoporphyrin. The presence of oxygen appears to be necessary for the formation of this polar porphyrin.
The effect of tropomyosin on the adenosine triphosphatase activity of desensitized actomyosin
Authors: Schaub M. C. , Perry S V , Hartshorne D. J.
Published: August 2015

1. Tropomyosin preparations of the Bailey type, and those prepared in the presence of dithiothreitol to prevent oxidation of protein thiol groups, inhibit the Ca2+-activated adenosine triphosphatase (ATPase) of desensitized actomyosin by up to 60%. 2. The inhibitory activity of myofibrillar extracts and tropomyosin survives various agents known to denature proteins but to the action of which tropomyosin is unusually stable, namely heating at 100° and mild tryptic digestion. It is destroyed by prolonged treatment with trypsin. 3. The ethylenedioxybis-(ethyleneamino)tetra-acetic acid (EGTA)-sensitizing factor present in extracts of natural actomyosin and myofibrils could be selectively destroyed, leaving unchanged the inhibitory effect on the Ca2+-activated ATPase. There was no correlation between the EGTA-sensitizing and the Ca2+-activated inhibitory activities of tropomyosin prepared under different conditions. 4. Optimum inhibition was achieved when tropomyosin and the myosin of desensitized actomyosin were present in approximately equimolar proportions. Tropomyosin had no effect on the Ca2+-activated ATPase of myosin measured under similar conditions. 5. Evidence is presented showing that the tropomyosin binds to desensitized actomyosin under the conditions in which the ATPase is inhibited.
An octadecatrienoic acid from Lamium purpureum L. seed oil containing 5,6-allenic and trans-16-olefinic unsaturation
Authors: Mikolajczak K. L. , Rogers Mary F. , Smith C. R. , Wolff I. A.
Published: August 2015

1. Lamium purpureum L. (Labiatae) seed oil contains 16% of a new acid characterized as (−)-octadeca-5,6-trans-16-trienoic acid (proposed trivial name ‘lamenallenic acid’) (Ia). The acid was isolated as its methyl ester by countercurrent distribution by using a combination of recycle–single withdrawal techniques. Methyl lamenallenate (Ib) is strongly laevorotatory. 2. The structure was established by infrared spectroscopy, nuclear-magnetic-resonance spectroscopy, quantitative hydrogenation and oxidative cleavage data of the original acid and of hydrazine partial reduction products. 3. Other unsaturated esters identified by their cleavage products were oleate, linoleate and linolenate. 4. A very small amount (less than 1%) of methyl laballenate [(−)-methyl octadeca-5,6-dienoate] was also isolated and identified.
The identification of prostaglandins E2, F2α and A2 from rabbit kidney medulla
Authors: Lee J. B. , Crowshaw K. , Takman B. H. , Attrep Katherine A. , Gougoutas J. Z.
Published: August 2015

Rabbit kidney medulla (10kg.) was homogenized in 5mm-disodium hydrogen phosphate and deproteinized with ethanol, and the concentrated supernatant solution was extracted at pH8 with light petroleum and at pH2 with chloroform. The acidic lipids present in the chloroform phase were separated on silicic acid columns into three biologically active fractions. The first fraction contained only vasodepressor activity; the second fraction contained both vasodepressor and non-vascular-smooth-muscle-stimulating activity; the third fraction contained both vasopressor and non-vascular-smooth-muscle-stimulating activity. Purification of each fraction by reversed-phase partition and thick-layer chromatography yielded three pure acids. Thin-layer chromatographic, spectroscopic and mass-spectral analysis of the acids and their methyl esters established their structures as prostaglandins E2, F2α and A2. Evidence is presented demonstrating that part or all of the prostaglandin A2 is formed during the isolation procedures from endogenous prostaglandin E2.
Biliary excretion of foreign compounds. Benzene and its derivatives in the rat
Authors: Abou-el-Makarem M M , Millburn P , Smith R L , Williams R T
Published: August 2015

1. The extent of the excretion in the bile of the rat of benzene and 21 of its simple derivatives was studied. 2. Some 16 compounds of molecular weight less than 200, and including neutral molecules (benzene and toluene), aromatic acids, aromatic amines and phenols, were injected in solution intraperitoneally into biliary-cannulated rats. Metabolites in the bile were identified and estimated. The extent of biliary excretion of these compounds was low, i.e. 0–10% of the dose in 24hr., and most appeared in the bile mainly as conjugates. 3. The biliary excretion of six conjugates of molecular weight less than 300, including three glycine conjugates, one sulphate conjugate, one glucuronic acid conjugate and two acetyl derivatives, was low (less than 3% of the dose). 4. It is concluded that simple benzene derivatives of molecular weight less than about 300 are poorly excreted in rat bile.
Metabolism of the reserve polysaccharide of Streptococcus mitis. Some properties of a pullulanase
Authors: Walker Gwen J.
Published: August 2015

1. A pullulanase has been separated from cell extracts of Streptococcus mitis. The enzyme was freed from transglucosylase by fractionation with ammonium sulphate. 2. Pullulanase was produced in the absence of inducers, and addition of glucose or maltose to the broth did not increase the yield of enzyme. 3. The pullulanase acted rapidly on α-(1→6)-bonds in substrates having the structure α-maltodextrinyl-(1→6)-maltodextrin, but had no action on isomaltose, 6-α-glucosylmaltodextrins or 6-α-maltodextrinylglucoses. 4. 6-α-Maltotriosylmaltodextrins were hydrolysed over 10 times faster than 6-α-maltosylmaltodextrins. 5. The branch linkages of amylopectin phosphorylase limit dextrin, glycogen phosphorylase limit dextrin and glycogen β-amylase limit dextrin were hydrolysed. The action of pullulanase on amylopectin and glycogen was accompanied by a rise in the iodine stain of 50% and 30% respectively. 6. A reversal of pullulanase action occurred on incubation with high concentrations of maltotriose. Condensation of maltosyl units to form a branched tetrasaccharide occurred less readily. 7. S. mitis pullulanase was rapidly inactivated at temperatures higher than 40°, and the enzyme did not recover activity on storage at room temperature.
Inhibition of staphylococcal α-toxin. The effect of aromatic polysulphonic acids on the lethal effect of α-toxin in mice
Authors: Arbuthnott J. P. , Lominski I. R. W. , Wright Margaret Robson
Published: August 2015

1. Certain aromatic polysulphonic acids, previously tested for inhibition of the haemolytic activity of staphylococcal α-toxin, together with some additional related compounds, were tested as possible inhibitors of α-toxin in mice. 2. Compounds that inhibited the haemolytic activity of α-toxin at concentrations of 0·16mm or less [compounds (I), (II), (IV), (V), (VII) and (VIII)] were found to inhibit the lethal effect of α-toxin. 3. With the exception of compound (VIII), amounts of 1mg. were required to inhibit 4 LD50 of toxin when the test compounds were premixed with α-toxin before injection; comparable inhibition with 0·3mg. of compound (VIII) was achieved without prolonged premixing. 4. Mixtures of α-toxin and compounds (I) and (II) containing an excess of test compound showed markedly diminished inhibitory activities. 5. The ‘half-molecule’ analogues of group 1 [compounds (III) and (XVIII)] were non-inhibitory. 6. Compounds (I)–(V), when administered separately from α-toxin by the same route (intraperitoneal), were active only when injected almost simultaneously with toxin, whereas compounds (VII) and (VIII) were strikingly inhibitory when injected 15min. before or after the toxin. 7. Compound (VIII) failed to inhibit the lethal effect of α-toxin when injected by a different route (intravenous).
Some properties of galactokinase in developing rat liver
Authors: Walker D G , Khan H. H.
Published: August 2015

1. The nature of the galactokinase present in the livers of foetal, newborn and adult rats was examined by the application of several separation procedures and by measurement of a range of kinetic parameters. 2. No evidence of enzyme heterogeneity at any stage of development was found during gel filtration on Sephadex G-100, column chromatography on DEAE-cellulose or a variety of electrophoretic procedures. 3. The Km values, inhibition characteristics and other kinetic parameters appear to remain constant during development. 4. Rat liver galactokinase activity does not adapt to dietary changes in either the adult or the newborn rat; hence it is unlikely that the presence of galactose in milk controls the enzymic activity profile during development. 5. On the present evidence it is concluded that only one form of galactokinase is present in rat liver and that the enzymic activity is controlled by non-dietary factors.
Effect of denervation on the synthesis of ribonucleic acid and deoxyribonucleic acid in rat diaphragm muscle
Authors: Manchester K L , Harris E J
Published: August 2015

1. The effect of unilateral denervation of rat diaphragm muscle on its content of nucleic acids and their incorporation of precursors was investigated. 2. After denervation the paralysed hemidiaphragm hypertrophies and within 3 days its content of RNA increases considerably. The concentration of DNA/unit mass remains fairly constant. 3. During this period there is some increase in the rate of incorporation of [14C]adenine into RNA, whereas there is some diminution in the rate of incorporation of [14C]orotic acid. 4. Incorporation of [14C]adenine and [3H]thymidine into DNA is much increased in the paralysed tissue, reaching its maximum by about the third day, but returning to normal by the tenth. 5. The significance of these results in relation to the hypertrophy after denervation is discussed.
The reversible inactivation of pig kidney alkaline phosphatase at low pH
Authors: Butterworth P. J.
Published: August 2015

1. Pig kidney alkaline phosphatase is inactivated by treatment with acid at 0°. 2. Inactivated enzyme can be partially reactivated by incubation at 30° in neutral or alkaline buffer. The amount of reactivation that occurs depends on the degree of acid treatment; enzyme that has been inactivated below pH3·3 shows very little reactivation. 3. Studies of the kinetics of reactivation indicate that the process is greatly accelerated by increasing temperature and proceeds by a unimolecular mechanism. The reactivated enzyme has electrophoretic and gel-filtration properties identical with those of non-treated enzyme. 4. The results can be best explained by assuming that a lowering of the pH causes a reversible conformational change of the alkaline phosphatase molecule to a form that is no longer enzymically active but is very susceptible to permanent denaturation by prolonged acid treatment. A reactivation mechanism involving sub-unit recombination seems unlikely.