Cooperative binding of primycin and gramicidin on erythrocyte membranes. A cation transport study

In this paper the authors present a comparative study of the actions of the antibiotics primycin and gramicidin on the erythrocyte membrane permeability. It has been found that both antibiotics have a nonlinear effect on the membrane permeability. Above a threshold antibiotic concentration, which is characteristic of the type of the antibiotic, the cation permeability of the erythrocyte membranes increases sharply. In the range of nonlinearity the transport-kinetic curves level off before achieving the equilibrium radioactive ion distribution between the extra- and intracellular spaces. A stochastic model of the cooperative and aspecific incorporation of antibiotic molecules into the membrane explains the experimental findings. The authors conclude that membrane permeability increases at the places where two or more antibiotic molecules form aggregates in the membrane.     

Aliphatic hydrocarbons obtained from iron carbides: Possible stellar and meteoritic implications

Origins of Life and Evolution of Biospheres -     

Fluorescent staphylococci as microbeads

Fixed bacteria of the protein A-rich Cowan I Staphylococcus strain were labelled with fluorescein isothiocyanate and used as the second-step reagent in an indirect immunofluorescent assay of specific cell-surface antigen expression. The results are documented with fluorescence microscopy and flow cytometry.     

Interaction between the Ca2(+)-ATPase and the proteolipid in artificial membranes

The interaction between the Ca2+ transport ATPase and the proteolipid of rabbit sarcoplasmic reticulum was analyzed by fluorescence energy transfer, using the following donor: acceptor combinations: Ca2(+)-ATPase tryptophan----IAEDANS-proteolipid; IAEDANS-ATPase----IAF-proteolipid; IAEDANS-proteolipid----IAF-ATPase. The observed energy transfer may indicate weak interaction between the Ca2(+)-ATPase and proteolipid, but collisional energy transfer definitely contributes. The energy transfer was abolished by deoxycholate or sodium dodecylsulfate at concentrations sufficient to solubilize the membrane. In view of the low proteolipid content of sarcoplasmic reticulum and the weak interaction suggested by the energy transfer, at best only a small fraction of ATPase molecules could exist in the form of ATPase-proteolipid complexes.     

Defensive behaviour in the pond snail, Lymnaea stagnalis: the whole body withdrawal associated with exsanguination

The freshwater snail L. stagnalis is known to be able to respond to a strong, noxious stimulus with a full retraction of the foot and head into the shell accompanied with expelling the blood through the hemal pore. We have found that this behavioural response, besides graded local retractions, can be triggered by mild tactile stimulation provided that the stimulus is repeated several times. Only a complete exsanguination could be obtained, indicating that it is an all-or-none defensive behaviour. In an electrophysiological investigation of isolated brain, a number of similarities were found between this all-or-none behaviour and the so-called input 3 to central neurons, as described by Benjamin and Winlow. These include ability to be selectively activated by high calcium solutions, and blocked by keeping the snails in a spoiled water. Injection of snails with naloxone (0.5-2.0/micrograms/g) or ergotamine (0.4/microgram/g) blocked selectively the whole body withdrawal induced by tactile stimulation, but not that induced by injection of a high calcium saline or acetylcholine solution, indicating that enkephalinergic and/or dopaminergic mechanosensory neurons might be involved. The consideration of available data has led to a working hypothesis that the activity of input 3 might be the neurophysiological correlate of the high threshold all-or-none whole body withdrawal associated with exsanguination.     

Cellular regulation of ADP-ribosylation of proteins. III. Selective augmentation of in vitro ADP-ribosylation of histone H3 in murine thymic cells after in vivo emetine treatment

Thymic cells were isolated at intervals of between 0 and 144 h from mice that received one intraperitoneal injection of emetine (33 mg/kg), and thymus weight, incorporation of [14C]leucine into proteins and [3H]thymidine into DNA in intact thymic cells, as well as initial rates of protein ADP-ribosylation in permeabilized cells [A. Sóoki-Tóth, F. Asghari, E. Kirsten, and E. Kun (1987) Exp. Cell Res. 170, 93] were simultaneously monitored. The effect of emetine as an inhibitor of protein synthesis [F. Antoni, N. G. Luat, I. Csuka, I. Oláh, A. Sóoki-Tóth, and G. Bánfalvi (1987) Int. J. Immunopharmacol. 9, 333] corresponds to the induction of sequential cellular events, such as cell exit and remigration, by other antimitotic agents [C. Penit and F. Vasseur (1988) J. Immunol. 140, 3315] and produces an activation of proliferation of cells reentering into this organ. Proliferation, as demonstrated by a large increase in DNA synthesis and entrance into S phase, was kinetically related to an apparent increase in poly(ADP-ribose) polymerase activity in thymic cells and a highly significant in vitro ADP-ribosylation of histone H3. Since no DNA fragmentation occurred in thymic cells, as tested by a fluorometric technique [C. Birnboim and J. J. Jevac (1981) Cancer Res. 41, 1889], it is probable that a selective activation of poly(ADP-ribose) polymerase may have been induced in cells that undergo differentiation and proliferation while repopulating the thymus.     

Immunocytochemical study of parvalbumin fibers and cell bodies in the rat hipothalamus

The distribution of parvalbumin (PA) cell bodies and fibers in the hypothalamus of the rat was studied using a monoclonal antibody and the avidin-biotin-peroxidase method. The densest clusters of immunoreactive perikarya were observed in the nuclei mamillare medialis, arcuatus and dorsomedialis hypothalami, whereas the corpus mamillare lateralis had the lowest density. The densest network of immunoreactive fibers was observed in the corpus mamillare lateralis and nucleus arcuatus. The corpus mamillare medialis contained a moderate number of PA fibers, whereas the nucleus dorsomedialis hypothalami had the lowest density of immunoreactive fibers. In addition, a large number of immunoreactive fibers was found in the tractus opticus and the tractus mamillo-thalamicus. Essentially, the distribution of PA in the rat hypothalamus after using a monoclonal antibody seems to be broader in comparison with previous studies carried out in the same diencephalic region of the rat. The presence of PA in several nuclei of the rat hypothalamus suggests that this protein could be directly or indirectly involved in neuroendocrine, limbic and visual functions.     

Catalytic activity of cytochrome oxidase and cytochrome c in apolar solvents containing phospholipids and low amounts of water

Cytochrome c and cytochrome oxidase, in bovine heart submitochondrial particles and in their purified forms, were transferred to a ternary system that contained phospholipids (10 mg/ml toluene), the apolar solvent toluene, and water at concentrations of 13-15 microliters (high water) and 3 microliters (low water) per milliliter of toluene. When the enzymes were transferred back to an all water system, they exhibited full catalytic capacity. In the low water ternary system, cytochrome c could be reduced by ascorbate introduced via inverted micelles. Also in this system, cytochrome oxidase was reduced by ascorbate and cytochrome c but its oxidation was highly impaired. Data on the kinetics of reduction by ascorbate of cytochrome c and cytochrome oxidase under these conditions are presented. Cytochrome oxidase reduced in the organic solvent by ascorbate failed to form a complex with CO, but formed a complex with cyanide introduced via inverted micelles. The oxidized and the ascorbate-reduced cytochrome oxidase-cyanide complex exhibited a trough at 415 nm and a peak at 433 nm. The extent and rate of formation of the cyanide complex were higher with the reduced form of cytochrome oxidase. To achieve protein-protein interactions (cytochrome c-cytochrome oxidase) in the ternary system, it was necessary to extract the two proteins together. There was no functional interaction when they were extracted separately and mixed. In the high water ternary system reduced cytochrome oxidase was not detected, and it oxidized ascorbate at a higher rate than in the low water system; however, this rate was several orders of magnitude lower than in aqueous media.     

Influence of the solvent on the conformational-dependent properties of random coil polypeptides. II. Mean square optical anisotropies and Kerr constants

Mean square optical anisotropies and molar Kerr constants were calculated for homopolypeptides of the 20 natural amino acids and of several enzymes and proteins in the random-coil state. The effect of hydration was taken into account in constructing the molecular potential that gives the conformational energies as a function of the rotational angles phi and psi of the backbone and chi(1) of the side chain. The Rotational Isomeric State model was used in calculated energies, the Valence Optical Scheme and the matrix calculus technique of Flory being employed in the evaluation of the optical properties. The results are compared with calculations for the same substances that were performed without taking into account the solvent, as well as with other similar studies. The Kerr constant is confirmed as being one of the most sensitive properties of a given polypeptide to the residue class and to the sequence of those residues.     

Stereological study of gonadotropes in the frog,Rana pipiens,after GnRH stimulation in vitro

Summary Previous physiological results have indicated the existence of two releasable pools of gonadotropins in amphibian pituitaries: an acute releasable pool that appears independent of protein synthesis, and a storage pool involved in chronic release that depends on protein synthesis. To elucidate the ultrastructural localization of these pools and the morphological changes induced in gonadotrope cells after treatment with gonadotropin-releasing hormone, we carried out a morphometric study of immuno-identified gonadotrope cells using an in vitro superfusion system. Treatment with gonadotropin-releasing hormone induced a degranulation of small (110–255 nm) and medium (236–360 nm) secretory granules as well as hypertrophy of the endoplasmic reticulum and Golgi complex. Simultaneous incubation with gonadotropin-releasing hormone and cycloheximide inhibited the release of secretory granules although the endoplasmic reticulum and Golgi complex were hypertrophied. These morphological results strongly suggest: (1) that gonadotropin-releasing hormone induces degranulation and hypertrophy of the biosynthetic machinery in gonadotrope cells; and (2) that the activation of the endoplasmic reticulum and Golgi complex by stimulation with gonadotropin-releasing hormone is independent of protein synthesis, while the release of secretory granules is protein synthesis-dependent. In addition, the second or storage pool of gonadotropin is associated mainly with the small and medium secretory granules.