Purification by preparative electrophoresis and characterization of histne H2B from ox pancreas

• 1.1. Highly purified histone H2B from ox pancreas has been isolated by preparative electrophoresis in polyacrylamide slab gel, at pH 2.7 using the fraction F2b as starting material.
• 2.2. This histone was characterized by amino acid analysis, end groups determination and the tryptic peptide maps.
• 3.3. Comparative studies of histone H2B from ox pancreas and homologous calf thymus histone show similarity of the amino acid compositions, amino terminal groups as well as the tryptic peptide maps.

     

Histone variants in diptera during metamorphosis

• 1.1. Patterns of histone variants from three diptera, Ceratitis capitata, Dacus oleae and Drosophila melanogaster were compared to mouse and to Plodia interpunctella variant patterns.
• 2.2. The three diptera contain histones which comigrate on two dimensional gels with H3.2, H3.3 and H4 in mouse and Plodia. H2A.1 and H2B.1 comigrate with Plodia H2A.1 and H2B.1 and are different from mouse, whilst H2A.Z has a different mobility to that of Plodia and mouse.
• 3.3. The iodinated peptides obtained from H2A.1 and H2A.2 of the diptera studied are compared to mouse and Plodia H2A.1 and H2A.Z peptides.
• 4.4. The histone variants from three developmental stages, larval, pupal and adult of the three diptera were identified and compared.
• 5.5. The same histone variant pattern is found through all stages of development.

     

Aggregation of the histone h1 from the fruit fly Ceratitis capitata through disulphide bridges. Studies on their complexes with DNA

• 1.1. Three different major components, BG1, BG2 and BG3, have been obtained when the histone H1 from the fruit fly Ceratitis capitata is oxidized in vitro, separated by gel permeation chromatography and characterized by both amino acid analysis and polyacrylamide electrophoresis.
• 2.2. BG1 and BG2 correspond to the aggregation products by formation of intermolecular disulphide bridges, while BG3 is a monomeric component which shows the presence of one intramolecular disulphide bridge.
• 3.3. Structural studies by both circular dichroism and controlled tryptic digestion on BG1, BG2 and BG3 show the native conformation of H1 from the insect changes slightly and in a different range in each component.
• 4.4. All the subfractions induce PSI structure in DNA and stabilize the double helix of DNA, although quantitative differences appear.

     

The occurrence and properties of histone H1° in quiescent rabbit tissue

• 1.1. The issue of the existence and the properties of histone H1° in quiescent rabbit tissues was approached by electrophoretic, Chromatographic and immunochemical techniques.
• 2.2. The results demonstrated that the rabbit tissues did contain a typical histone H1°, its properties being comparable to those of H1° from all other sources studied thus far.

     

A comparative study of the basic nuclear proteins from sperm of bivalve molluscs

• 1.1. Basic nuclear proteins from spermatozoa of the three mollusc species belonging to the class Bivalvia have been analyzed using one- and two-dimensional electrophoresis.
• 2.2. Four nuclear basic proteins have been purified and their amino acid compositions determined.
• 3.3. In these spermatozoa histone-type proteins coexist with protamine-like proteins.
• 4.4. The protamine-like proteins that have been studied show different electrophoretic behavior but in general are similar, with a high content of lysine, arginine, alanine and serine.
• 5.5. Interspecific variability has been found for the H1-like histone.

     

Chasing Tails: Cathepsin-L Improves Structural Analysis of Histones by HX-MS,

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Highlights

• •Cathepsin-L is introduced as a novel protease for HX-MS studies.
• •Cathepsin-L improves resolution of traditionally challenging histone tails.
• •Cathepsin-L can be readily combined with pepsin for improved protein coverage.
• •In-solution dynamics of the H3.1 and H4 monomers reveal extensive EX1 kinetics.

     

Adaptative features of amazon fishes: Hemoglobins,hematology, intraerythrocytic phosphates and whole blood Bohr effect of Pterygoplichthys multiradiatus (Siluriformes)

• 1.1. Hemoglobin, hematological parameters, intraerythrocytic phosphates and whole blood Bohr effect of Pterygoplichthys multiradiatus, from the Amazon river, were studied in three different conditions: in their natural environment, acclimated to normoxia and acclimated hypoxia conditions.
• 2.2. Nine anodal hemoglobin fractions were detected on starch gel electrophoresis. No qualitative differences in the Hb electrophoretic patterns were detected in the three studied groups.
• 3.3. Hematocrit, hemoglobin concentration, MCV, MCHC and MCH were different among studied conditions.
• 4.4. GTP was almost absent in the blood of animals in natural conditions and acclimated to hypoxia, but was present at a concentration similar to ATP in normoxic acclimated animals.
• 5.5. There is a tendency for higher Hb-O₂ affinity for hypoxic acclimated/acclimatized animals.

     

Variations in the lactate dehydrogenase activity during the development of the shrimp Palaemon serratus

• 1.1. The lactate dehydrogenase (LDH) from Palaemon serratus muscle has been studied throughout the development of the animal.
• 2.2. Enzymatic activities have been traced by polyacrylamide gel electrophoresis and kinetic studies.
• 3.3. The existence of two enzymes (L₁ and L₂) has been demonstrated.
• 4.4. During the larval development, both L₁ and L₂ remain at a low level.
• 5.5. After the larvae hatch L₁ and L₂ gradually rise although L₁ is predominant.
• 6.6. Measurement of kinetic parameters shows that the general behaviour of the enzymes of the embryo resembles that of the adult enzymes.
• 7.7. However, one can observe during the development a constant increase in the affinity of the enzyme towards its substrate, lactate.

     

Sodium transport in red blood cells of lamprey LAmpetra fluviatilis

• 1.1. Unidirectional Na⁺ influx in lamprey red blood cells was determined using ²²Na as a tracer.
• 2.2. Total Na⁺ uptake and amiloride-inhibitable Na⁺ influx increased in a saturable fashion as a function of external Na⁺ concentration (Na_e).
• 3.3. At 141 mM Na_e, the average value of net Na⁺ influx was 13 ± 1.1 and the amiloride-sensitive Na⁺ influx was 5.3±1.1 mmol/l cells per hr (±SE).
• 4.4. The amiloride-sensitive component of Na⁺ influx was significantly activated by 10⁻⁵ M isoproterenol, by 2 × 10⁻⁵ M DNP, and by cell shrinkage.
• 5.5. Furosemide (1 mM) had no effect on the Na⁺ transport in red cells.
• 6.6. The residual amiloride-insensitive component of Na⁺ transport was a linear function of Na_e in the range of 5–141 mM. This transport seems to be accounted for by simple diffusion.

     

Age-dependent proteins in eyes of annual killifishes Pterolebias longipinnis detected by 2-dimensional electrophoresis

• 1.1. Eye proteins of Pterolebias longipinnis have been analyzed by 2-dimensional isoelectric focusing SDS-polyacrylamide gel electrophoresis during aging from adolescence until normal death.
• 2.2. The protein pattern on the gels changed gradually with progressing age.
• 3.3. In senescent eyes, three protein spots appeared for a time and 36 disappeared from the pattern.
• 4.4. The isoelectric points of the proteins in the presence of urea and the molecular weights in an unreducing buffer are presented.

     

Lipid components and utilization in consumers of a seagrass community: An indication of carbon source

• 1.1. The lipid components of three animals, the rock crab Nectocarcinus integrifons, the rock flathead Platycephalus laevigatus and the southern garfish Hyporhamphus melanochir, feeding in the seagrass beds at Corner Inlet, Victoria, Australia have been examined in detail in order to provide further information on seagrass community structure.
• 2.2. Biological marker compounds detected within animal gut content material were used to recognize dietary sources and then utilized by community members.
• 3.3. Both H. melanochir and N. integrifons have been shown to ingest and to varying degrees incorporate seagrass lipid material, thus further confirming the importance of seagrass carbon in the Corner Inlet environment.
• 4.4. The southern sea garfish H. melanochir is observed to remove C₁₈ PUFAs (polyunsaturated fatty acids) from ingested seagrass material.
• 5.5. Seagrass sterols are altered during incorporation into the lipids of this fish.
• 6.6. Lipid-rich digestive juices play a role in the digestive processes of all three animals.
• 7.7. Components tentatively identified as (NMI) (non-methylene interrupted) fatty acids have been detected in the lipids of the garfish H. melanochir and the crab N. integrifons.
• 8.8. The fecal material of all three animals represent possible sources of these lipids (NMI acids) in Corner Inlet sediments.
• 9.9. Based on lipid compositional data, N. integrifons feeds on Posidonia australis detritus and associated epiphyte material.
• 10.10. The removal of both plant and epibiota cellular lipids along the digestive tract of the crab was observed, although structural components such as long chain mono- and α,ω-dicarboxylic acids, which have been previously recognized as seagrass marker lipids are not directly absorbed.

     

Isolation and characterization of phospholipase A2, from Agkistrodon bilineatus (common cantil) venom

• 1.1. Phospholipase A₂ was isolated from Agkistrodon bilineatus venom by Sephadex G-75 and CM-Cellulose column chromatographies.
• 2.2. The purified phospholipase A₂-I gave a single band on disc polyacrylamide gel electrophoresis, isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis.
• 3.3. The enzyme preparation had a molecular weight of 14,000, isoelectric point of pH 8.77 and possessed 123 amino acid residues.
• 4.4. The purified phospholipase A₂ possessed lethal, indirect hemolytic and anticoagulant activities.
• 5.5. The enzyme hydrolyzed the phospholipids phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl inositol (PI) and phosphatidyl serine (PS).
• 6.6. The concentration of mouse diaphragm was inhibited and the contraction of guinea pig left atrium was increased by phospholipase A₂-I.
• 7.7. Phospholipase A₂ activity of this preparation was inhibited by ethylenediamine tetraacetic acid, p-bromo phenacyl bromide, n-bromo succinimide or dithiothreitol, but not by diisopropyl fluorophosphate or benzamidine.

     

Some observations on the behaviour of the sodium efflux in barnacle muscle fibres towards lithium ions

• 1.1. The behaviour of the Na efflux towards Li⁺ was studied using single barnacle muscle fibres as a preparation.
• 2.2. It is found that the Na efflux into Li⁺-ASW (artificial seawater) is reduced and that this effect is not fully reversed by returning back to Na⁺-ASW.
• 3.3. Preinjection of 100 mM-EGTA reduces the magnitude of the fall of the Na efflux into Li⁺-ASW.
• 4.4(a). The remaining Na efflux into Li⁺-ASW is further reduced by external application of 10⁻⁴ M-ouabain. (b) The remaining Na efflux in ouabain-poisoned fibres is reduced by replacing Na_e by Li⁺. However, some fibres show a rise rather than a fall.
• 5.5. Fibres loaded with NaCl (by injection) show a prompt and sustained stimulation of the Na efflux when Na_e is replaced by Li⁺. A similar but less pronounced response is often seen with ouabain-poisoned fibres.
• 6.6. Injection of LiCl (e.g. a 2 M-solution), causes a 20% fall in Na efflux. Subsequent replacement of Na_e by Li⁺ fails to bring about a fall in the remaining efflux.
• 7.7. Itis concluded that the Na efflux in these fibres consists of a Na-Na exchange diffusion component which is not mediated by the Na-K pump and that its operation is interrupted by injecting Li⁺. The relative size of this component is about one-fifth and not one-half of the Na efflux.

     

Cardiac responses of larval and adult tiger salamanders to submergence and emergence

• 1.1. The ECG of aquatic Amhystoma tigrinum from the Colorado Rocky Mountains was recorded while the animals submerged and emerged in water. Older larvae and metamorphosed adults were compared.
• 2.2. Free-swimming animals of both types showed slight emergence tachycardia when taking a “gulp” of air.
• 3.3. Preventing access to air for 30 min or more resulted in a slight bradycardia in larvae. Some adults responded with increased, others with decreased, heart rate depending on their level of excitement.
• 4.4. Restraining the animals before forced submergence caused a greater bradycardia than when unrestrained.
• 5.5. Low dissolved oxygen accentuated the cardiac responses of larvae to submergence but not in adults.
• 6.6. Atropine only partially blocked the diving responses of both forms.
• 7.7. The degree of submergence bradycardia seems to be a function of the ability to extract oxygen from water. It probably is not an adaptation to diving in these forms. Instead the submerged heart rate in these predominantly aquatic salamanders may be the “normal” rate with emergence tachycardias for breaths of air.

     

Dynamics of Hlo content in rat liver after partial hepatectomy

• 1.1. The changes in the lysine-rich histone subfraction hl₀ have been quantitatively studied in rat liver during the regeneration period after partial hepatectomy.
• 2.2. A gradual decrease in this protein was found early after operation with a minimal value around the time of maximal mitotic activity.
• 3.3. The reduction in the hl₀ content paralleled well the increasing number 0f cells in the cell cycle.

     

NAD(P)H dehydrogenase from rabbit and rat liver: Purification and some properties

• 1.1. NAD(P)H dehydrogenase from rabbit liver was purified to electrophoretic homogeneity using a procedure also found applicable for the rat liver enzyme.
• 2.2. Rabbit and rat liver enzymes showed different behaviour in isoelectric focusing and different K_m values and turnover numbers.
• 3.3. Both enzymes were inhibited to similar extents by warfarin.
• 4.4. The rabbit enzyme is composed of two subunits of mol. wt 27,000 and contained 1 FAD group per subunit.
• 5.5. Some absorption and circular dichroism properties of the rat enzyme are shown.

     

Localization and characterization of hemoproteins in the central nervous tissue of some gastropods

• 1.1. The red coloured ganglia of Lymnaea stagnalis and Planorbarius corneus, and the colourless ganglia of Helix pomatia and Cepaea nemoralis have been investigated by the benzidine reaction for the characterization and localization of hemoproteins.
• 2.2. For L. stagnalis and P. corneus the reaction deposits appear only within glial cells, whereas for the central nervous tissue of H. pomatia and C. nemoralis the hemoproteins are constituent of cytosomes within neurons only.
• 3.3. By gel electrophoresis these neuro-hemoproteins of L. stagnalis and P. corneus show similar electrophoretical mobility and differ from the hemoproteins in the hemolymphs respectively.
• 4.4. Experiments with time depending cultured ganglia prove the important part of the neuro-hemoproteins in the gas exchange between the hemolymph and the avascular ganglia.
• 5.5. The connection of the environmental conditions of the animals studied here with regard to the occurrence of hemoglobin-like proteins and carotinoids in the ganglia was discussed.

     

Main serum protein of rainbow trout (Salmo gairdnerii): its biological properties and significance

• 1.1. Main serum α₁-protein (α₁P) of rainbow trout was purified and its biochemical and physico-pathological properties were studied.
• 2.2. α₁P was suggested to be a primitive protein having both properties of albumin and AFP in serum proteins of mammals according to the following results.
• 3.3. Molecular weight (75,000), two kinds of molecules (pI 4.55 and 5.05) and amino acid composition.
• 4.4. Dye- or ConA binding activity.
• 5.5. Estrogen binding activity and inhibitory effect on lymphoblastoid-forming activity.
• 6.6. Possible osmotic regulator.
• 7.7. Significant elevation of blood α₁P level in the course of hepatoma induction.

     

Oxygen consumption of the sandy-beach whelk Bullia digitalis meuschen at different levels of activity

• 1.1. O₂ consumption of suspended Bullia digitalis is not related to water current speed or degree of turbulence, where these are kept constant.
• 2.2. The highest levels of O₂ uptake at 15°C are obtained by producing fluctuating surges of turbulence, the animals responding to changes in the movement of water.
• 3.3. In buried animals O₂ consumption decreases with time in the absence of water movements.
• 4.4. Burrowing and surface crawling require less energy than transport in the surf.