Homology between ricin and Ricinus communie agglutinin: Amino terminal sequence analysis and protein synthesis inhibition studies

The existence of three forms of ricin and two forms of the Ricinus communis agglutinin (RCA) was established using cation exchange chromatography, isoelectric focusing, and polyacrylamide gel electrophoresis. The preparation of the RCA we obtained was 60–75 times more potent than ricin in the agglutination of erythrocytes, but was about 4% as effective as an inhibitor of cell-free protein synthesis. When reduced with 2-mercaptoethanol, the RCA was activated 3000-fold as an inhibitor of cell-free protein synthesis, whereas ricin was activated about 600-fold by the same treatment. A mixture of the RCA A chains was about one-fifth as effective as the ricin A chain in the inhibition of cell-free protein synthesis. The purified polypeptide subunits of the castor bean lectins were subjected to automated Edman degradation. The sequence for 17 of the first 19 residues of the agglutinin A chain was determined. The first seven residues of the ricin A chain were determined and they are identical with those of the RCA A chain. Nineteen turns of Edman degradation on the RCA B chain resulted in the identification of 18 amino acids. The sequence determined for the first 17 residues of the ricin B chain was identical with that of the RCA B chain. It is likely that the identity of the ricin/RCA A and B chain sequences extends further along the polypeptide chains than the sequences we have determined. The similar structural and catalytic potentials of the RCA and ricin suggest that they bear a precursor-product relationship.     

Control of acetylcholine receptor mobility and distribution in cultured muscle membranes. A fluorescence study

The molecular control of the distribution and motion of acetylcholine receptors in the plasma membrane of developing rat myotubes in primary cell culture was investigated by fluorescence techniques. Acetylcholine receptors were marked with tetramethylrhodamine-labeled α-bungarotoxin and lateral molecular motion in the membrane was measured by the fluorescence photobleaching recovery technique. Three types of experiments are discussed: (I) The effect of enzymatic cleavages, drugs, cross-linkers, and physiological alterations on the lateral motion of acetylcholine receptors and on the characteristic distribution of acetylcholine receptors into patch and diffuse areas. (II) Observation of the distribution and/or motion of fluorescence-labeled concanavalin A receptors, lipid probes, cell surface protein, and stained cholinesterase in acetylcholine receptor patch and diffuse areas. (III) The effect of a protein synthesis inhibitor and electrical stimulation on membrane incorporation of new acetylcholine receptors.Some of the main conclusions are: (a) acetylcholine receptor lateral motion is inhibited by concanavalin A plant lectin and by anti-α-bungarotoxin antibody, but marginally enhanced by treatment with a local anesthetic; (b) patches are stabilized by an immobile cellular structure consisting of molecules other than the acetylcholine receptors themselves; (c) this structure is highly selective for acetylcholine receptors and not for other cell membrane components; (d) acetylcholine receptor patch integrity and diffuse area motion are independent of direct metabolic energy requirements and are sensitive to electrical excitation of myotube; (e) lipid molecules can move laterally in both acetylcholine receptor patches and diffuse areas; and (f) acetylcholine receptor lateral motion in diffuse areas and immobility in patch areas are not altered by specific agents which are known to affect extrinsic cell surface proteins, or cytoplasmic microfilaments and microtubules.     

The wavelength dependence of the turbidity of solutions of macromolecules

The experimental parameter describing the wavelength dependence of the turbidity of solutions of macromolecules is usually the negative slope of the graph of the logarithm of optical density vs the logarithm of the in vacuo wavelength, λ₀. Such slopes are the apparent exponents of 1/λ₀ in the turbidity equation. Their values depend upon the way the destructive interference of the scattered light, the refractive index increment, and the solvent refractive index change with wavelength. In this study, expressions for the wavelength exponents for isotropic rods, spheres, and random coils have been obtained and evaluated by representing the intraparticle interference functions, Q(λ₀), with series in even powers of 1/λ₀ and the refractive properties with Cauchy relations. Comparisons of calculated and observed exponents at wavelengths in the visible spectral region for aqueous solutions of four viruses have been made: for R17, T7, and PM2 bacteriophage, the exponents are greater than four; whereas for tobacco mosaic virus, they are less than four. The application of the turbidity relations to determine the size and molecular weight of biological macromolecules is discussed.     

Réflexions critiques sur l'écologieet la systématique des Lingules actuelles et fossiles

New topics on ecology and systematics of recent and fossil Lingulids lead to an obvious revision of our knowledges on this zoological group. At first, the recent species need systematics and taxonomy on the bases of new described specific criteria (as, morphology of deltidial areas, muscle disposition); the results are briefly indicated. But, in fossil species, disorder and disparity of used characteristics are emphasized.The general conceiving on ecology of Lingulids are reviewed and discussed, especially on bathymetry and salinity; sediment and oxygenation conditions; taphocoenosis and lingulid «communities. On recent species, all these points are also studied, especially some ecological requirements (salinity, bathymetry, grain size), and mechanism of burrowing ability, burrow living positions in the sediments, as shell preservations after death and fossilization, to facilitate the paleobiotope interpretations. Recent animals are euryhalin, surviving at salinities from about 13 to 42‰; they could be considered as well adapted to waters with strong salinity fluctuations. They show preference to fine sand bottoms (lowest particle size about 40–60 μm). Their bathymetric distribution occurs between 0 and about 500 m (Lingula especially between 5–50 m; Glottidia 15–70 m). The isotherms 8–10°C seem to restrict their geographical and bathymetric distribution.Therefore, some post-palaeozoic lingulid bedsare studied or redescribed on the bases of the above discussed characteristics, and new interpretations on the environmental situation are given (Trias of Vosges Mountains; Oligocene from Japan; Eocene of London Basin). More caution must be used in study of fossil Lingulids that are not especially animals living in infralittoral bottoms with low salinity and deficient oxygenation, as generally accepted.     

Nuclear magnetic resonance study of hydrogen-bonded ring protons in oligonucleotide helices involving classical and nonclassical base pairs.

A study of the exchangeable ring nitrogen protons in aqueous solutions of oligonucleotide complexes involving Watson-Crick base pairs as well as Hoogsteen pairs and other nonclassical hydrogen bonding schemes shows that resolvable resonances in the low-field (-10 to -16 ppm from sodium 4,4-dimethyl-4-silapentanesulfonate) region can be detected in a variety of structures other than double stranded helices. Ring nitrogen proton resonances arising from the following hydrogen-bonding situations are reported: (1) AT and GC Watson-Crick base pairs in a self-complementary octanucleotide, dApApApGpCpTpTpT; (2) U-A-U base triples in complexes between oligo-U15 and AMP; (3) C-G-C+ base triples in complexes between oligo-C17 and GMP at acid pH; (4) s4U-A-s4U base triples in complexes between oligo-s4U15 and AMP, all of which involve both Watson-Crick and Hoogsteen base pairing to form triplexes; (5) C-C+ base pairing between protonated and unprotonated C residues in oligo-C17 at acid pH; and (6) I4 base quadruples in the four strand association among oligo-I at high salt. The behavior of the dA3G-CT3 helix is consistent with both fraying of the terminal base pairs and presence of intermediate states as the helix opens. In the monomer-oligomer complexes, under the conditions used here, the exchange appears to be governed by the dissociation rate of monomer from the complex. These findings suggest that those tertiary structure hydrogen bonds in tRNA involving ring nitrogen protons should have representative resonances in the low-field (11-16 ppm) proton NMR region in H2O.     

Systematic relationships in the narrow-leaved species of chenopodium of the Western United States

Brittonia - In a study of ten taxa ofChenopodium occurring primarily in the western United States and designated as the narrow-leaved complex, data from morphology, flavonoid chemistry, and somatic...     

Interactions of lectins with (Na+ + K+)-ATPase of eel electric organ.

Interaction of lectins with a detergent-solubilized ATPase from eel electric organ was studied. Concanavalin A, which binds to alpha-mannosides, altered the rate of enzyme migration in agar and inhibited the formation of an antigen-antibody precipitate: other lectins had no such effects. Concanavalin A similar amounts partially inhibited (Na+ + K+)-ATPase; this inhibition was reversible by alpha-methylglucoside. There was no corresponding effect of concanavalin A on the potassium p-nitrophenylphosphatase. Concanavalin A also did not interfere with ouabain binding. Thus, concanavalin A binds to an antigenic region also involved in Na+ and/or ATP binding, but does not interact with a K+ site.