Collagen diversity in the sea urchin,strongylocentrotus purpuratus

• 1.1. Pepsin insensitive fragments of collalgen extracted from tube feet and peristome have α 1 and α 2 bands that differ in apparent molecular weights from each other and from human type 1 collagen.
• 2.2. A monoclonal antibody that reacts with the ξ I band and a low molecular weight fragment of tube foot collagen does not react with either peristome collagen or human type I collagen.
• 3.3. Measurements of the axial periodicity of native fibers of tube foot and peristone collagens indicate they have D values that differ significantly from each other and from reported values of vertebrate type I collagen.
• 4.4. We propose that there are diverse and specialized types of collagen in sea urchins that are heterogeneously distributed in the extracellular matrix of different tissue.

     

Phosphorylation of a 25,000-dalton protein in slow and fast muscles of the chicken

• 1.1. Protein phosphorylation in intact chicken latissimus dorsi muscle, slow anterior (ALD) and fast posterior (PLD), was compared.
• 2.2. A major difference in [³²P]phosphate incorporation was found between the ALD and PLD in a 25,000-dalton heat soluble protein.
• 3.3. The 25,000-dalton protein was purified from both the ALD and PLD.
• 4.4. The two proteins had similar amino acid composition and both contained approximately 1 mole phosphate per mole of protein.
• 5.5. The difference in their content of radioactive phosphate was determined to be due to faster turnover in the ALD.

     

Microheterogeneity of melanosome-bound tyrosinase from the harding-passey murine melanoma

• 1.1. This study was directed towards the characterization of the origin of the microheterogeneity displayed by mammalian tyrosinase, the enzyme responsible for pigmentation in mammals.
• 2.2. Tyrosinase was purified from the Harding Passey murine melanoma, fractionated into a continuous series of subisozymic forms, and analyzed using various chemical and immunological probes.
• 3.3. Treatment with neuraminidase revealed that all the forms had similar amounts of sialic acid, and reactivity with various carbohydrate specific lectins showed that the isozymes also contained subterminal galactose, N-acetylglucosamine, and mannose, but lacked α-fucose.
• 4.4. Amino acid composition data indicated that the polypeptides of all the forms had identical residue contents.
• 5.5. The sum of the evidence further supports the theory that the isozymic forms demonstrable for mammalian tyrosinase represent intermediate processing stages of the enzyme from the nascent protein chain to the fully glycosylated, high molecular weight form of tyrosinase that is localized within melanin granules.

     

Identification and estimation of dopamine released from the anterior byssus retractor muscle of Mytilus californianus in response to nerve stimulation

• 1.1. Small quantities of sea water were recyclically perfused over the surface of paired anterior byssus retractor muscles of Mytilus californianus.
• 2.2. Dopamine was identified in the perfusate by thin-layer chromatography.
• 3.3. Stimulation of the pedal ganglion caused the dopamine content of the perfusate to increase.
• 4.4. A significant increment of release of dopamine was detected at stimulation frequencies above 3 Hz and increased progressively with increase in stimulation frequency.
• 5.5. The possibility of a role for dopamine as a relaxing or inhibitory neurotransmitter in Mytilus is considered in relation to the present and related evidence and to the actions of 5-HT, the probable relaxing transmitter.

     

Subcellular distribution of metals within the kidney of the bivalve Mercenaria mercenaria (L.)

• 1.1. The subcellular distribution of nine transition metals (plus four additional elements) was measured in the kidney tissue of the quahog, Mercenaria mercenaria.
• 2.2. Elemental analyses of the subcellular fractions indicated three main patterns of metal distribution within kidney cells.
• 3.3. Barium, iron, manganese and lead were associated primarily with kidney granules.
• 4.4. Cadmium, copper, potassium and magnesium were found mainly in the cytosolic fraction.
• 5.5. Calcium, phosphorus and zinc were found in all isolated fractions, probably reflecting the important roles that these elements play in bivalve metabolism.
• 6.6. The organelle composition of the isolated subcellular fractions was determined using marker enzyme assays and microscopic techniques.

     

Purification and characterization of a manganese containing superoxide dismutase from bovine heart mitochondria

• 1.1. A manganese containing superoxide dismutase from bovine heart mitochondria was isolated and characterized.
• 2.2. It has a molecular weight of about 86,000 and is composed of 4 noncovalently bound subunits of equal size.
• 3.It appears to contain 2 mole manganese per mole enzyme.
• 4.The carbohydrate content is very low.
• 5.The specific activity and amino acid composition are similar to those of other mitoehondrial superoxide dismutases.
• 6.The enzyme forms complexes with ampholytes and can therefore not be analysed by isoelectric focusing.

     

Immunochemical and immunohistochemical determination of ubiquitin in molluscan (Mytilus edulis) smooth muscle

• 1.1. Immunochemical and immunohistochemical distribution of ubiquitin in the anterior byssus retractor muscle (ABRM) of Mytilus edulis was investigated.
• 2.2. In immunostaining, specific ubiquitin immunoreactivity was observed in the cross-sectioned ABRM, and was uniformly localized in this section.
• 3.3. The amount of free ubiquitin in the ABRM homogenate was 130 ± 4.6 ng/mg protein by western blot analysis, and ubiquitin conjugates were found at about 25, 29 and 200–230 kDa.
• 4.4. These findings were similar to those obtained in the skeletal muscle of rat.

     

Characterization of procollagen type iv and of its heterogenous accumulation products in the extracellular matrix of bovine lens cells in culture

• 1.1. Bovine lens epithelial cells synthesized in culture medium type IV procollagen which can be labelled both with [³H]mannose and [¹⁴C]glucosamine.
• 2.2. This procollagen eluted on Agarose column ahead the component of collagen type I. After reduction and alkylation it has an apparent molecular weight of 180,000.
• 3.3. The behaviour of the pepsin extractible collagen from both culture medium and cell layer submitted to salf fractionation and thermal gelation was similar to that of type IV collagen isolated from lens capsule.
• 4.4. Ion exchange chromatography and CNBr peptides profiles of these collagen molecules are similar to that obtained with lens capsule collagen. However, some heterogeneity is detected in the CMC Chromatographic profile obtained with the collagen isolated from the cell layer.
• 5.5. Analysis on SDS PAGE of the collagen isolated from the cell layer gives an heterogenous profile similar to that of lens capsule collagen and contrasts with homogenous electrophoretic pattern of collagen extracted from culture medium.

     

The occurrence of relaxing granules in the muscle of the locust,Locust migratoria

• 1.1. A granular or vesicular fraction was isolated from the muscles of the locust, Locust migratoria, and allowed to react with isolated myofibrils in order to examine whether or not this fraction could act as a relaxing factor as in the case of mammalian muscle.
• 2.2. The granules obtained from the muscle of this insect were very effective in inhibiting myofibrillar ATPase, whether the myofibrils were prepared from the same muscles or from the rabbit muscles.
• 3.3. Superprecipitation of a puridied actomyosin preparation was greatly retarded by the addition of these granules.
• 4.4. Evidence was put forward that the granules are acting, in the presence of ATP, by removing calcium from myofibrils because of their strong calcium-binding capacity.
• 5.5. These observations seem to suggest that the insect granules are also capable of acting as relaxing factor and that all muscles, whether in the vertebrates or in the invertebrates, can be considered as identical systems in so far as the chemical mechanism of contraction-relaxation is concerned.

     

Comparative hematology: Studies on class aves,domestic turkey (meleagris gallopavo)

• 1.1. The mean clotting time (C.T.) for turkey blood was 71.0 min in glass and 83.0 min in siliconized tubes. Accelerated C.T.s with rabbit brain or partial reagent were over 60 sec but with turkey brain or RVV they were less than 20 sec.
• 2.2. Fibrinogen averaged 450 mg/dl. Coagulation factors, assayed in human systems, were very low with the exception of XIII and VIII (0.57 U/ml).
• 3.3. Thrombocytes aggregated during clotting and with collagen but not with ADP. Thrombocytes contained microtubules, an open canalicular system, a large nucleus but no α granules or dense bodies.
• 4.4. Hemostasis appeared dependent on an extrinsic pathway.

     

Effects of temperature and metabolic inhibitors on contraction of anterior byssus retractor muscle of Mytilus

• 1.1. Anterior byssus retractor muscle of Mytilus (ABRM) was stimulated to contract by ACh (acetylcholine) and effects of temperature (5–30°C), FDNB (1-fluoro 2,4 dinitro-benzene) and IAA (iodoacetic acid) on tension response were examined.
• 2.2. Isometric tension was highest at the temperature range of 10–20°C and decreased at higher and lower temperature than that range.
• 3.3. The rate of tension decay after washing of ACh was accelerated by the increase of temperature.
• 4.4. Tension redevelopment after release of 1 % during contraction was much smaller at 5°C than at 20°C.
• 5.5. Tension development by ACh and the rate of tension decay after washing of ACh were remarkably decreased by the treatment of FDNB or IAA.
• 6.6. The above results were discussed from the viewpoint that energy metabolism might be related to catch.

     

Amino acid composition and the protein form of procarboxypeptidase a purified from the pancreas of the sei whale Balaenoptera bolealis

• 1.1. Procarboxypeptidase (W-PCPA) was purified from the pancreas of the sei whale Balaenoptera bolealis.
• 2.2. W-PCPA was obtained as a homogeneous protein in polyacylamide gel disc electrophoresis.
• 3.3. W-PCPA has a molecular weight of 75,000.
• 4.4. Amino acid composition of W-PCPA was compared with that of bovine procarboxypeptidase as A S5 (PCPA-S5).
• 5.5. W-PCPA may be two subunits, and the aggregate form may resemble PCPA-S5.

     

Effect of metabolic inhibitors on hyperosmotically induced free amino acid accumulation in the isolated foot muscle of the brackish-water bivalve Corbicula japonica

• 1.1. Accumulation of free amino acids (FAA) in the isolated foot muscle of the brackish-water bivalve Corbicula japonica during the initial stage of hyperosmotic stress was quantitatively and qualitatively similar to that in the foot of the intact animal.
• 2.2. Aminooxyacetate (AOA), a transaminase inhibitor, markedly inhibited alanine accumulation and promoted ornithine accumulation in the isolated foot. Iodoacetate (IAA), a glycolytic inhibitor, caused no significant alteration in the alanine level and the TLC pattern of FAA. Both the inhibitors scarcely influenced the pool size of total ninhydrin positive substances (NPS).
• 3.3. A major part of the carbon of accumulated FAA during the initial stage of hyperosmotic stess did not seem to arise from glycolysis.
• 4.4. Free d-alanine as well as l-alanine accumulated in isolated foot muscle exposed to hyperosmotic stress.

     

Characterization of NaK ATPase from the gills of the freshwater prawn Macrobrachium rosenbergii (de Man)

• 1.1. The specific activity of Na-K ATPase was determined from the microsomal preparation of gills dissected from adult Macrobrachium rosenbergii.
• 2.2. Maximal ATPase activity was achieved at a substrate concentration of 0.5 mM ATP.
• 3.3. Optimal enzyme activity was obtained at pH of 7.5.
• 4.4. The Arrhenius plot of Na-K ATPase activity revealed a marked discontinuity at 30°C. “Mg” ATPase activity did not exhibit a marked discontinuity.
• 5.5. The E_a for Na-K ATPase and “Mg” ATPase was 14.6 kCal/mole and 9.31 kCal/mole respectively. Q₁₀ values for Na-K ATPase was 2.34 and for “Mg” ATPase 1.65.
• 6.6. ATPase activity and gill homogenate protein concentration exhibited a linear relationship up to 130 μg protein/ml.
• 7.7. Na-K ATPase activity was inhibited by 10⁻³ M ouabain. It was equally inhibited by the removal of K⁺ from the reaction medium.

     

Haematology of the australian eastern quoll,Dasyurus viverrinus

• 1.1. A variety of haematological parameters were determined in adult Dasyurus viverrinus.
• 2.2. Haemoglobin and red cell counts were high with a very low mean cell volume.
• 3.3. Basophils are absent but the eosinophils contain small numbers of basophilic granules which may indicate a dual role for this cell.
• 4.4. “Ring Form” leucocytes are present.
• 5.5. Three types of red cell picture could be identified, some animals showing large numbers of spherocytes, spicule cells, and inclusion bodies.
• 6.6. These cells resemble those found in some inherited human haemolytic anaemias but there was no evidence of haemolysis in the animals.
• 7.7. An alkali resistant haemoglobin component is present.

     

Composition of milk of the pygmy sperm whale (Kogia breviceps)

• 1.1. A 315cm lactating female pygmy sperm whale, Kogia breviceps, accompanied by a 156cm female calf stranded on south Miami Beach, Dade County, Florida, on 13 March 1974, were used in the experiment.
• 2.2. A sample of milk from the lactating female contained less fat and more lactose than most cetecean milks previously analyzed.
• 3.3. Palmitic and oleic acids predominated in Kogia milk fat and long chain unsaturated fatty acids were lacking.
• 4.4. Kogia milk protein contained several unidentified constituents.
• 5.5. The chlorinated hydrocarbon residues in the milk were (ppb, wet weight): total DDT—1670; dieldrin—37; PCB—1203.

     

The occurrence of brassicasterol and epibrassicasterol in the chromophycota

• 1.1. Sterols were identified from eight isolates of five species in the Chromophycota that were cultured axenically and harvested in the stationary phase.
• 2.2. Analyses were performed on four strains from the Prymnesiophyceae, two strains from the Cryptophyceae and one from the Bacillariophyceae. Most strains examined contained only one major sterol, 24-methyl-22-dehydrocholesterol.
• 3.3. Analysis by capillary GC, HPLC, and in one instance NMR, showed that the two strains provisionally identified as Isochrysis contained brassicasterol (24β-methyl-22-dehydrocholesterol); whereas, all other species examined contained primarily epibrassicasterol (24α-methyl-22-dehydrocholesterol).
• 4.4. Stigmasterol (24α-ethyl-22-dehydrocholesterol) accompanied epibrassicasterol in Pleurochrysis carterae.
• 5.5. Analyses of C-24 alkyl isomers in these algae may provide useful information concerning their taxonomic placement.
• 6.6. The occurrence of both isomers of 24-methyl-22-dehydrocholesterol in oysters is explained by the occurrence of both isomers among algae which are probably dietary sources for oysters.

     

Fluorescence studies on the interaction of muscle M-line proteins,creatine kinase and the 165,000 dalton component,with each other and with myosin and myosin subfragments

• 1.1. The binding of 8-anilino-l-naphthalene sulfonate (ANS) by M-line creatine kinase (CPK) and the 165,000 dalton protein component was studied by fluorescence.
• 2.2. One mole of ANS binds to a mole of each M-protein and the binding site on these two M-proteins is hydrophobic in nature.
• 3.3. M-line proteins labeled with ANS were used to demonstrate their interaction with myosin and myosin subfragments 1 and 2.
• 4.4. A unique finding of this study, that labeled M-line CPK binds to subfragment 2 of myosin, is of significance from a structural view point, since only the rod portions of myosin molecules are exposed at the M-line and therefore able to interact with CPK.
• 5.5. The use of a sulfhydryl fluorescent probe, N-(2-iodoacetyl)-N-(5-sulfo-l-napthyl) ethylene diamine, (1,5-AEDANS), has shown that the essential sulfhydryl group on CPK is very important for its interaction with both myosin and the 165,000 dalton M-line protein component.

     

Phospholipids composition and metabolism in the hemolymph of Carcinus maenas (Crustacea,Decapoda)—Effect of temperature

• 1.1. The distribution of different phospholipids and the repartition of fatty acids extracted from hemolymph of crab Carcinus maenas are analysed.
• 2.2. The action of the temperature on the lipid composition is also determined: an increase of content of PE and a slight rise of the degree of unsaturation of fatty acids are found at lower temperatures.
• 3.3. The specific radioactivity of total phospholipids, phosphatidyl-choline and phosphatidylethanolamine from hemolymph of Carcinus maenas is studied from two radioactive precursors (³²Phosphorus and ³H]ethanolamine). Results suggested that the conversion of PE into PC by methylation could take place in hepatopancreas of Carcinus maenas.
• 4.4. The specific radioactivity of phospholipids from these two same radioactive compounds is increased following a variation in the environmental temperature.
• 5.5. The composition of hemolymph lipids could be a direct reflection of the lipid metabolism of the hepatopancreas and that the temperature alters the rate of the phospholipid exchange between hepatopancreas and hemolymph.
• 6.6. It is suggested that these lipid alterations occur in order to permit crab Carcinus maenas to support large changes in environmental temperatures.

     

A thioredoxin from the extreme thermophilic Archaeon Sulfolobus solfataricus

• 1.1. A purification procedure for a thioredoxin from the extremophilic archaeon Sulfolobus solfataricus is described.
• 2.2. The thioredoxin is active in the dithiothreitol-dependent reduction of insulin disulfide bonds.
• 3.3. The thioredoxin is a monomer of 24,800 Da; it is an acidic protein with a pi of 4.5.
• 4.4. The protein is stable to heating for 3 hr at 90°C.
• 5.5. The amino acid composition of S. solfataricus thioredoxin is reported.