Volume 1, Issue 4
December 1979, pages 357-468
pp 357-367 December 1979
The aminoacylation of tRNA catalysed by valyl-tRNA synthetase (EC 184.108.40.206) and isoleucyl-tRNA synthetase (EC 220.127.116.11) fromMycobacterium smegmatis is dependent on the presence of divalent metal ions. Polyamines alone, in the absence of metal ions, do not bring about aminoacylation. In the presence of suboptimal concentrations of Mg2+, polyamines significantly stimulate the reaction. Of the cations tested, only Mn2+, Co2+ and Ca2+ can partially substitute for Mg2+ in aminoacylation, and spermine stimulates aminoacylation in the presence of these cations also. At neutral pH, spermine deacylates nonenzymatically aminoacyl tRNA. AMP and pyrophosphate-dependent enzymatic deacylation of aminoacyl-tRNA (reverse reaction) is also stimulated by spermine. The inhibitory effect of high concentration of KC1 on aminoacylation is counteracted, by spermine. The low level of activity between pH 8.5–9.0 at 1.2 mM Mg2+ is restored to normal level on the addition of spermine. The inhibitory effect of high pH on aminoacylation in the presence of low concentration of Mg2+ is also prevntedvby spemine.
pp 369-375 December 1979
The biosynthesis of fatty acids from [l-14C]-acetate in the chicken liver slicesin vitro was inhibited by cAMP, adenosine, 5′-AMP, 3′-AMP, ATP, NAD and FAD but not by adenine, guanine or inosine. The minimum structural requirement for inhibition appears to be adenosine. The inhibitory action of adenosine, 5′-AMP and NAD on fatty acid synthesis is likely to be mediated by adenosine or its metabolites since adenosine deaminase reverses the inhibition while it has no effect on the inhibition by cAMP; thus, the inhibitory effect of cAMP is probably not mediated through its hydrolysis products, 5′-AMP, or adenosine.
pp 377-383 December 1979
The photodecomposition of bilirubin follows first order kinetics with akB value of 12.5 × 10-3 min-1. In the presence of a model system generating superoxide anions, such as xanthine-xanthine oxidase, thekB value was 103 × 10-3 min-1 This ten-fold enhancement ofkB value by xanthine-xanthine oxidase was abolished when the reaction mixture was supplemented with a superoxide ion scavenger— superoxide dismustase. Further, known singlet oxygen quenchers like Β-carotene and bistidine did not prevent the enhancement of bilirubin oxidation by xanthine-xanthine oxidase, thereby ruling out the obligatory conversion of Superoxide anion to singlet oxygen. It is concluded that radical oxygen mediated bilirubin degradation might be a natural catabolic route for the bile pigment degradation during oxygen stress.
pp 385-392 December 1979
The results presented in this paper show that lysis of human erythrocytes by linoleic acid is not caused by peroxidation of the fatty acid. Peroxidase, superoxide dismutase and scavengers of O2− and OH had no effect on the lysis while catalase showed only marginal inhibition suggesting that O2−, OH, O2− and H2O2 do not play any direct role in hemolysis by linoleic acid. Generators of H2O2 inhibited the lysis completely and methemoglobin cells were more resistant to hemolysis by linoleic acid. The fatty acid did neither bind to nor fomed complex with red cell ghosts. Membrane oxidation of sulphydryl groups was also not involved in the lysis. Β-Carotene, retinol and bile salts enhanced the lysis, while, cholesterol but not cholesterol acetate, inhibited it. Taurocholate-pretreated cells were more susceptible to linoleic acid lysis. These observations suggested-that lysis by linoleic acid may be due to its detergent property.
pp 393-400 December 1979
Proteolytic activity was detected in neem (Azadirachta indica) exudate gum when tested with casein and albumin as substrates. The enzyme activity was separated into two fractions by chromatography on TEAE-cellulose after EDTA treatment. Both the enzyme fractions were fairly stable to high temperatures and wide range of pH conditions. The pH optima were found to be around 6.5. Phenylmethyl sulphonylfluoride inhibited the activity of both the fractions. EDTA, Β-mercaptoethanol, tosylamide phenylethylchloromethylketone, tosyllysine chloroimethylketone,p-chloromercuribenzoate and dithiobis-2-nitrobenzoie acid did not affect the activity of the two enzyme fractions. The two fractions had no hydrolytic action on a variety of synthetic substrates tested.
pp 401-407 December 1979
A simple procedure for the isolation of cathepsin-B from bovine pancreas employing ammonium sulphate fractionation, DEAE cellulose chromatography and Sephadex G-200 gel filtration is described. The purified enzyme gave a single band on polyacrylamide gel electrophoresis. The molecular weight as determined by gel filtration of the enzyme was 26,850. ItsKm andVmax values were 12.8 mM and 0.303 Μmol/min/mg, respectively. TheKi for iodoacetamide was 0.16 mM.
pp 409-424 December 1979
Five commercial preparations of glucoamylases (three fromAspergillus niger, one each fromAspergillus foetidus andAspergillus candidus) were purified by ultrafiltration, Sepharose-gel filtration and DEAE-sephadex chromatography. Two forms of the enzyme, namely glucoamylase I and glucoamylase II were obtained from the fungi except from one strain ofA. Niger. All the enzymes appeared homogeneous by electrophoresis and ultracentrifugation. The specific activities varied between 85 and 142 units. The pH and temperature optima were between 4 and 5, and 60‡C respectively. The molecular weight as determined by the sodium dodecyl sulphate-polyacrylamide gel electrophoresis ranged from 75,000 to 79,000 for glucoamylase I and 60,000 to 72,000 for glucoamylase II. OnlyA. niger glucoamylases contained phenylalanine at the N-terminal end. The amino acid composition of the enzymes was generally similar. However,A. niger andA. foetidus glucoamylases, in contrast toA. candidus enzymes, contained greater percentage of acidic than of basic amino acids. The enzymes contained 15 to 30% carbohydrate and 49 to 57 residues of monosaccharides per mol.A. niger enzymes contained mannose, glucose, galactose, xylose and glucosamine but theA. candidus enzyme lacked xylose and glucose and only xylose was absent inA, foetidus enzymes. Majority of the carbohydrate moieties were O-glycosidically linked through mannose to the hydroxyl groups of seline and threonine of the polypeptide chain.
pp 427-432 December 1979
Mitochondria isolated from the gill tissues of the fishSarothredon mossambica were analysed for their macromolecular content, following transfer from freshwater to media of higher salinity. The results suggest a breakdown of mitochondria during the initial phases of the stress and a regeneration during continued exposure. Also all the synthetic machineries, in general, seem to be triggered in gill tissue during continuous exposure to hyperosmotic media.
pp 433-439 December 1979
Uptake of Co2+ by cobalt-resistant strain is dependent on Co2+ concentration in the medium and is linear with time. The uptake is unaffected by metabolic inhibitors and decreased at low pH values. The uptake is independent of temperature in the range 0–40‡ C. The transport system is a passive diffusion process, unlike in the parent wild type strain where it is energy-dependent. It is possible that Mg2+ transport system is not involved in Co2+ transport in this strain, since the Co2+ uptake is not suppressed by Mg2+ as in the parent strain.
pp 441-446 December 1979
Exposure of the silkworm(Philosamia ricini) larvae to cold temperature (2‡C) and subsequent exposure to room temperature (29‡ C) resulted in the mortality of the larvae.
Cold exposure brought about significant decrease in enzymic activity of proteases, aminotransferases, diacylglycerol lipase and in the amounts of some haemolymph sugars and polyols. However, glycerol increased sharply in response to severe cold exposure. There was also a marked increase in the levels of protein, pyruvate, total free amino acids, total lipid, phospholipid and triacylglycerols.
In the colder environment, carbohydrates served as the energy source. Glycerol appears to have conferred cryoprotection to the cold-stressedPhilosamia ricini larvae.
pp 447-455 December 1979
Extracellular polypeptides released by both N2-grown [peptide I] and NO3-grown [peptide II]Anabaena L-31 have molecular weight of approximately 3,500 but have distinctly different amino acid composition. Acid hydrolysis of the peptide I fraction (obtained by separation on Sephadex G-25) yielded ten amino acids whereas that from peptide II fraction yielded only 3 amino acids. On addition to a freshly inoculated N2-grown culture, the peptide I fraction stimulated pro-heterocyst and to a lesser extent heterocyst differentiation, whereas the peptide II fraction strongly inhibited differentiation. The inhibitory effect of polypeptide II fraction could not be relieved by methionine sulphoximine, which by itself enhances differentiation, but was greatly relieved by addition of the peptide I fraction. The data suggest but does not prove, thatAnabaena L-31 synthesises “inducer” or “inhibitor” peptides which could possibly control pattern formation.
pp 457-465 December 1979
Corpora lutea removed from pregnant hamster deprived of endogenous luteinizing hormone for varying periods were compared for their responsiveness to externally added luteinizing hormone. The corpora lutea removed on the 8th day of pregnancy exhibited a dose-dependent increase in progesterone production in response to added luteinizing hormone upto a concentration of 2.5 Μg/ml. The total progesterone synthesised by the corpora lutea decreased with increase in the duration ofin vivo luteinizing hormone deprivation. However, the hormone deprivation had to be for a minimum period of 24 h before a marked reduction in thein vitro responsiveness could be seen. Neutralisation of endogenous luteinizing hormone increased the luteal cholesterol ester concentration, whilein vitro incubation of such tissue with luteinizing hormone resulted in a marked reduction in cholesterol ester levels. Corpora lutea removed from hamsters on day 8, 15 and 16 of pregnancy when compared for their responsivenessin vitro to added luteinizing hormone showed that the luteal tissue of day 8 produced more progesterone relative to those of day 15/16. In contrast, depletion of free and esterified cholesterol increased with the increase in age of corpora lutea (from 15% on day 8 to 67% on day 16).
pp 467-468 December 1979