Cotranslational fatty acylation of mucus glycoprotein. Addition of palmitic acid to peptidyl-tRNA occurs prior to peptide chain completion and its release

• 1.1. The fatty acylation of mucus glycoprotein nascent peptides was investigated using [³H]palmitic acid and [³⁵S]methionine-labeled peptidyl-tRNA of rat gastric mucous cells.
• 2.2. The mucus glycoprotein peptidyl-tRNA fraction was found to contain covalently bound palmitic acid in its complexes.
• 3.3. RNase digestion of the mucus glycoprotein peptidyl-tRNA released [³H]palmitic acid labeled peptides which, on SDS-polyacrylamide gel, separated into a multitude of bands ranging in size from 2000 to 60,000 Da.
• 4.4. The analyses of low molecular weight peptides revealed that palmitic acid was present in methionine-labeled peptides containing 30–43 amino acids and those of 18–25 amino acids or larger devoid of methionine, but was not identified in methionine-labeled peptides containing 10–15 amino acids.
• 5.5. The results indicate that the N-terminal fatty acylation of mucus glycoprotein nascent peptides is a cotranslational process which is occuring in an immediate vicinity of the signal peptide fragment.

     

Fatty acid profile of the digestive gland and mantle in the bivalve Macoma balthica

• 1.1. Digestive gland and mantle fatty acids were studied in spring and summer in the bivalve Macoma balthica off the southern coast of Finland. The presence of lipids was also examined histochemically in various clam tissues.
• 2.2. the neutral lipid content of the digestive gland increased ca 4.5-fold during the annual growth period.
• 3.3. Neutral lipid fatty acids of the digestive gland, of which palmitoleic, eicosapentaenoic and palmitic acids were predominant, were clearly distinguished from phospho- and glycolipid fatty acids.
• 4.4. The degree of unsaturation of phospholipid fatty acids was higher in the cold season both in the digestive gland and mantle, mainly due to the titer of eicosapentaenoic acid.

     

Spingophosphonolipids in microsomes of Diplodon delodontus. Distribution and effect on the membrane microviscosity

• 1.1. The distribution of ceramide aminoethylphosphonate (CAEP) in microsomal membranes obtained from different tissues of the bivalve mollusc Diplodon delodontus was determined.
• 2.2. The concentration of CAEP reached from 9 to 19% of the total microsomal polar lipids, depending on the kind of tissue.
• 3.3. Palmitic acid was the main fatty acid in the ceramide moiety, followed by stearic and eicosamonoenoic acids.
• 4.4. Artificial membranes were prepared with microsomal phospholipids or phospholipids plus sterols, with and without the addition of CAEP.
• 5.5. It was shown that the phosphonate confers minor mobility to the membranes. This effect is more effective when the membrane contains the natural sterols and the phospholipids are unsaturated.

     

Influence of dietary sources of fat on lipid synthesis in mink (Mustela vison) mammary tissue

• 1.1. The fatty acid composition of the triglyceride fraction of mink milk sampled during mid-lactation (day 28 post partum) from two nursing mink was compared to that of plasma samples and to the fatty acid composition of the feed rations used.
• 2.2. Chemical analysis of the triglyceride composition of mink milk demonstrated only minute concentrations of fatty acids with a chain length below C₁₄.
• 3.3. The saturated C_16:0- and C_18:0-unit fatty acids in mink milk made up for 24–40% of the total amount of fatty acids extracted, the remainder being represented by mono and polyunsaturated long-chain (C₁₆-C₂₄) fatty acids.
• 4.4. Preliminary in vitro experiments proved the incorporation of¹⁴C-labelled glucose, acetate or palmitate into triacylglycerols in cultures of mink mammary tissue to be linear for at least 2 hr.
• 5.5. The in vitro capacity for de novo fatty acid synthesis in mink mammary tissue using 14C-labelled glucose or acetate was low, i.e. ranging from 0.096–0.109 nmol/g (fresh tissue)/min, and amounted to only about 5% of that obtained in the case of [¹⁴C]palmitic acid incubation.
• 6.6. Following ¹⁴C-labeIled acetic or palmitic acid incubation of mink mammary tissue neither desaturation nor chain elongation was observed.
• 7.7. In response to long-term feeding on rations with two different sources of animal fat (F = fish oil or L = lard) the influence of compositional changes in dietary neutral lipids on the fatty acid composition of the lipids of mink milk is discussed.

     

Human placental atp-diphosphohydrolase: Biochemical characterization,regulation and function

• 1.1. Kinetic and physico-chemical studies on human placental microsomal fraction confirmed that the ATPase and ADPase activities detected in this fraction correspond to the enzyme ATP-diphosphohydrolase or apyrase (EC 3.6.1.5). These include substrate specificity, and coincident M_r and pI values of both ATPase-ADPase activities.
• 2.2. This enzyme hydrolyses both the free unprotonated and cation-nucleotide complex, the catalytic efficiency for the latter being considerably higher.
• 3.3. Microsomal apyrase is insensitive to ouabain and Ap5A. The highly purified enzyme was only inhibited by o-vanadate, DBS and slightly by DCCD.
• 4.4. Apyrase seems to be a glycoprotein from its interaction with Concanavalin-A.
• 5.5. Preliminary studies on the essential amino acid residues suggest the participation of Arg, Lys and His residues, and discard the requirement of −SH, COO⁻, −OH, and probably also Tyr and Trp.
• 6.6. Two kinetic modulatory proteins of apyrase were detected in placental tissue. An activating protein was found in the soluble fraction and an inhibitory protein was loosely bound to the membranes.
• 7.7. The proposed in vivo function for apyrase is related to the inhibition of platelet aggregation due to its ADPase activity, which is supported by the direct effect on washed platelets and by its plasma membrane localization.

     

Proteolysis post mortem in North Atlantic krill

• 1.1. The autoproteolytic processes in selected species of North Atlantic krill, Meganyctiphanes norvegica (M. Sars), Thysanoessa inermis (Krøyer) and T. raschii (M. Sars) have been examined at 0°C by following the release of peptides and free amino acids.
• 2.2. The krill contains high levels of peptide hydrolases, and autoproteolysis seems to be due mainly to digestive enzymes localized in the hepatopancreas and the intestinal tract of the animals.
• 3.3. During autoproteolysis the individual amino acids were generally released at rates corresponding to their proportion in the bulk protein of the krill. The major exceptions were alanine which accumulated in amounts larger than was to be expected from the composition of the krill protein, and glutamic acid/glutamine, aspartic acid/asparagine, arginine, and to some extent glycine, proline and serine, which accumulated to a lesser extent than was to be expected.
• 4.4. Storage of krill for 1 week resulted in only minor changes in the total content of amino acids as determined after acid hydrolysis, with the exception of alanine which increased in concentration.
• 5.5. The results suggest that the formation of free alanine is partly due to reactions other than proteolysis.
• 6.6. The release of free amino acids was accompanied by a considerable increase in the amount of small peptides, and glutamic acid/glutamine, aspartic acid/asparagine, glycine and proline tended to accumulate in these peptides.
• 7.7. The autoproteolytic activity of the Thysanoessa species showed seasonal variations, probably in response to food availability. In the case M. norvegica, the results suggest that there are smaller fluctuations in the level of proteolytic enzymes, probably indicating less pronounced variations in the food intake over the year.

     

Lipid digestion in cod (gadus morhua)

• 1.1. Cod, 2.6–3.4 kg. were fed a mixed diet of sprat, capelin oil and wheat flour.
• 2.2. Lipids from the feed, stomach and four intestinal segments were separated into tri-, di- and monoglycerides and free fatty acids and analysed by GLC.
• 3.3. All lipolytic products were concentrated in 14:0, 16:0 and 18:0, up to 60% and extremely low in the ω-3 fatty acids.
• 4.4. Residual triglycerides contained 80% of saturated and monoenoic fatty acids.
• 5.5. Linoleic acid increased from 2% in feed TG to 10% in TG of the rectum.

     

Hemocyanin in Oniscus asellus (Isopoda)

• 1.1. An immunological crossreactivity, as demonstrated by Western blotting, exists between O. asellus hemocyanin and anti-keyhole limpet (Megathura crenulata) hemocyanin.
• 2.2. Using the anti-keyhole limpet hemocyanin, the presence of hemocyanin was detected in the hepatopancreas.
• 3.3. The amino acid composition of the hemocyanin of O. asellus was found to be similar to that of other isopods.

     

Glycogen phosphorylase in lobster hepatopancreas: action of proteases on the enzyme

• 1.1. In lobster hepatopancreas, extracellular protreases cause the inactivation of glycogen phosphorylase.
• 2.2. The proteolysis of glycogen phosphorylase purified from rabbit muscle by these proteases has been shown by SDS-polyacrylamide gel electrophoresis.
• 3.3. A cell isolation technique has allowed us to remove proteases of extracellular digestion and to measure glycogen phosphorylase activity in lobster hepatopancreas.
• 4.4. The glycogen phosphorylase activity seems to be mainly associated with R cells while it could not be detected in B cells.

     

Seasonal variation in the energy metabolism in an estuarine crab,Chasmagnathus granulata (Dana, 1851)

• 1.1. The effects of seasonal variation on the carbohydrate and lipid metabolism of the Chasmagnathus granulata were investigated.
• 2.2. Glycemia is high in winter and summer and low in spring and fall.
• 3.3. The glycogen content in the hepatopancreas and muscle is higher in fall and winter, and decreases during spring and summer.
• 4.4. The muscle lipids are higher in summer, and decrease during fall and winter whereas hepatopancreas lipids are higher except in the fall.
• 5.5. The crabs show change in the metabolic pattern of lipids and carbohydrates during the seasons of the year.

     

Lipid composition of three geographical sources of brine shrimp nauplii (Artemia sp.)

• 1.1. Percentage of triacylglycerols (TG), free fatty acids (FFA) and phospholipids (PL) in the total lipids, the fatty acid composition of each of these lipid classes, and the percentage of cholesterol were determined by gas chromatography in three geographical sources (San Francisco Bay, SFB; Chinese, CH; Colombian, COL) of brine shrim (Artemia sp.) nauplii.
• 2.2. There were no significant differences among sources of brine shrimp in total lipids, TG or FFA with means for all sources of 17.8, 65.8 and 10.9%, respectively. Percentage of phospholipid was significantly higher in SFB and CH sources of brine shrimp, 25.1 and 26.5%, respectively, than in COL 18.3%.
• 3.3. Marked differences in percentages of 18:3 (n-3) (linolenic acid) and 20:5 (n-3) (eicosapentaenoic acid or EPA) were found among brine shrimp sources, and concentration of these two fatty acids were usually inversely related within sources. The CH source contained higher concentrations of EPA ( > 9.0%) than the COL and SFB sources (< 5.0%) in all three lipid classes analyzed. No 22:6 (n-3) (docosahexaenoic acid or DHA) was found in any brine shrimp source.
• 4.4. Fatty acid compositions of the TG and PL were similar and did not differ among sources of brine shrimp, while the FFA had a lower percentage of polyunsaturated fatty acids, but was similar among sources of brine shrimp.
• 5.5. Differences in n-3 fatty acid composition indicated a difference in nutritional quality among sources of brine shrimp for feeding larval fish.

     

Changes of the fatty acid composition in smolts of masu salmon (Oncorhynchus masou), associated with desmoltification and sea-water transfer

• 1.1. Tissue lipid compositions of desmoltified yearlings of masu salmon (Oncorhynchus masou) obtained by keeping smoltified fish in fresh water, were examined and compared to those of smoltified fish before and after transfer to sea-water (SW).
• 2.2. Lipid contents of muscle, liver, gut and gills of desmolts tended to increase compared to those of initial smolts.
• 3.3. The increased proportion of triacylglycerol (TG) and decreased proportion of phospholipids (PL) characterized the tissue lipids of desmolts.
• 4.4. Liver and muscle lipids showed no distinct differences both in content and proportion between initial and SW smolts, but gut and gill lipids of SW smolts decreased in content accompanied by a decrease of TG and an increase of PL in proportion.
• 5.5. Excepting muscle non-polar lipids, tissue lipids of desmolts contained more mono-unsaturated fatty acids and saturated fatty acids and less polyunsaturated fatty acids (PUFA), especially (n-3) PUFA such as 22:6(n-3), than those of initial and SW smolts.
• 6.6. No large differences in fatty acid composition were seen between initial and SW smolts except for the gut.
• 7.7. The proportion of (n-3) PUFA in the gut of SW smolts was higher than that of initial smolts.
• 8.8. The results indicated that masu salmon smolts can modify their lipid metabolism to adapt to ambient salinity changes. The proportion of (n-3) PUFA particularly in polar lipids, or in osmoregulatory organs such as gut and gills, was seen to be critical in lipid types of freshwater- or sea-water-adapted fish.

     

Lipid composition of the red and green forms of Actina equinia from the black sea

• 1.1. The fatty-acid composition of the red and green forms of Actinia equina from the Black Sea have been determined by methods involving silver-ion HPLC and GC-MS.
• 2.2. The fatty acid compositions of both forms of A. equina resemble those of most marine invertebrates. Substantial amounts of C₂₀ and C₂₂ polyunsaturated fatty acids were found.
• 3.3. Balck Sea Actenia equina contains a large amount of plasmalogens, mainly phospotidylcholine and phosphatidylserine plasmalogens.
• 4.4. The red form of A. equina contains more arachidonic acid in glycolipid fraction and more phosphatidylethanolamine and its plasmalogen, while the green from contains more sphingomyelin. These differences are an indication that the species A. equina can be divided into two subspecies—green and red.

     

Effects of milk fat,unhydrogenated and partially hydrogenated vegetable oils on serum lipoproteins in growing pigs

• 1.1. Crossbred Yorkshire (Yorkshire × Landrace) pigs were fed butter oil, cream, low erucic acid rapeseed oil, sunflower oil and partially hydrogenated sunflower oil in amounts representing 30% of energy for periods of up to 13 weeks.
• 2.2. After 13 wk of feeding serum total cholesterol levels of pigs fed milk fat were significantly higher than of pigs fed vegetable oils.
• 3.3. The difference in cholesterol was mainly due to an increase in the density range of 1.063–1.125 g/ml containing pig LDL₂ and some HDL.
• 4.4. A shift towards smaller LDL particle size was apparent in pigs fed milk fat.
• 5.5. The effects of dietary trans fatty acids did not differ from cis polyunsaturated or monounsaturated fatty acids.

     

Ionic dependence and vanadate sensitivity of (Na+, K+)-ATPase isolated from branchial sac of ascidians

• 1.1. Ion dependence and vanadium-induced inhibition on branchial sac ATPase in five species of ascidian Phlebobranchiata (vanadium-accumulating) and Stolidobranchiata (iron-accumulating) were studied.
• 2.2. The ATPase was obtained from the microsomal fraction, which was prepared from each ascidian branchial sac.
• 3.3. The ATPase was dependent on Mg²⁺ and activated by exogenous Na⁺ + K⁺.
• 4.4. Ouabain inhibited the ATPase activity in vitro, 10 μM to 100 μM vanadate, in vitro, suppressed the (Na⁺, K⁺)-ATPase.

     

Comparative study of the endemic freshwater fauna of lake baikal—I. Phospholipid and fatty acid composition of two mollusc species,Baicalia oviformus and Benedictia baicalensis

• 1.1. Lipid and phospholipid compositions of endemic freshwater molluscs belonging to the class Gastropoda, Baicalia oviformus and Benedictia baicalensis, were studied.
• 2.2. The fatty acids composition of total lipids, neutral, glyco- and phospholipid fraction was investigated by capillary gas chromatography-mass spectrometry.
• 3.3. Ninety-five fatty acids were identified: 23 saturated (both iso- and anteiso-), 28 monoenoic, 14 dienoic and 30 polyenoic.
• 4.4. High percentage of the two main acids, 18:4 and 18:4(n-3) in phospholipid and glycolipid fractions were identified.
• 5.5. A number of unusual polyunsaturated fatty acids, such as 19:4, 18:5(n-3), 24:4(n-6), 24:5(n-6), 24:6(n-3), and furanoid acids, were found.

     

Lipoxygenase of Thermoactinomyces vulgaris,purification and characterization of reaction products

• 1.1. A lipoxygenase preparation was obtained from Thermoactinomyces vulgaris and was purified by affinity chromatography on a linoleyl aminoethyl sepharose column.
• 2.2. Two active fractions were obtained.
• 3.3. The fraction obtained by elution with 100 mM borate buffer pH 9.0 was used in the subsequent work.
• 4.4. Th. vulgaris lipoxygenase oxidized linoleic acid into two products: 13-HPOD and 9-HPOD at a ratio of 44 to 56, respectively.
• 5.5. The identification and characterization of the isomers was done by HPLC, I.R. and mass spectrometry.
• 6.6. When arachidonic acid was used as substrate, 15-HPETE and 15-HETE were found to be the main enzymatic products.

     

The digestive proteases of langostilla (pleuroncodes planipes,decapoda): their partial characterization,and the effect of feed on their composition

• 1.1. Three methods of recuperating and preserving enzyme activity from freshly-caught langostilla were assessed. In the pressing and acetone extract methods, the recovered specific activity was similar.
• 2.2. Protease activity was higher between 6.5 and 8 pH, and was sensitive to high temperatures.
• 3.3. In PAGE and serine inhibition assays, one fraction resembled bovine trypsin.
• 4.4. The composition of proteins and molecules bearing protease activity from the hepatopancreas and stomach of both fed and starved animals was similar, indicating proteases are not induced but constitutive.

     

Sex-specific gene expression in distinct tissues of the shrimp Penaeus vannamei

• 1.1. Soluble proteins extracted from male and female Penaeus vannamei tissues such as eyes, eyestalks, brain, nerve cord, hemolymph, heart, muscle, hepatopancreas, hepatopancreas membrane and cuticular epidermis were analyzed and compared by high-resolution mini-two-dimensional polyacrylamide gel electrophoresis (mini-2D-PAGE).
• 2.2. In each shrimp tissue a large number of discrete polypeptides was observed.
• 3.3. The polypeptide patterns from the same tissue of female and male shrimp were mostly similar but both qualitative and quantitative differences were noted, suggesting the presence of sex-specific gene products in various shrimp tissues.
• 4.4. Future applications of these results are discussed.

     

Phosphatase activity in the hepatopancreas of Helix aspersa

• 1.1. Phosphatase acid (PhA) activity in the digestive gland (hepatopancreas) of the common garden snail Helix aspersa has been investigated using cytochemical methods.
• 2.2. All the cells composing this gland show PhA activity, the distribution pattern differing according to the cell type.
• 3.3. The digestive cells show the most widely distributed reaction product (brush border, phagolysosomes, multivesicular bodies and autophagic vacuoles).
• 4.4. In the excretory cells this activity appears in large sacs, while in the calcium cells the reaction product is abundant in the calcium granules.
• 5.5. Cellular digestion processes performed by each of these cell types is discussed together with their role in the detoxification of heavy elements derived from the environment.