Effect of LiCl on compression and tension properties ofPophyra perforata tissue

The mechanical properties of various differentiated regions of thePorphyra perforata thallus and the effect of LiCl were studied by performing compression and tension tests. Among the various differentiated tissues, the holdfast area was high in its compressive modulus of elasticity and tensile modulus of elasticity, possibly related to its thick matrix. Vegetative non-dividing tissue and vegetative dividing tissue were the most flexible and strong, showing the highest percentage elongation at break and tensile strength. The patch area, which is a transition zone leading to sexually mature tissue, had moderate values of tensile properties. Meanwhile, sexually differentiated male and female tissues had the highest compressive modulus of elasticity and lowest tensile properties. Thes tisues tended to crumble easily. Treatments in LiCl, as used for DNA extraction, resulted in a decrease in both compressive modulus of elasticity (87%) and tensile modulus of elasticity (54%). After treatment of tissue for chromosome staining in a method using LiCl, there was a marked decrease in tensile modulus of elasticity (49%), while the compressive modulus of elasticity remained unchanged. Such mechanical changes verify the softening effect of LiCl on the seaweedP. perforata tissue.Author for correspondence     

Digestion of seaweeds by the marine amoeba Trichosphaerium

A crude enzyme preparation from the marine amoeba Trichosphaerium was used to produce protoplasts from Sargassum muticum, Macrocystis pyrifera Porphyra perforata, and other red and brown marcroalgae. Cortical and medullary protoplasts of Sargassum, which were impossible to obtain using mixtures of previously available enzymes have now been prepared. Intact inner cortical and medullary protoplasts of Macrocystis, which were not observed in past isolations were obtained. Improved protoplast yields of as much as 500 fold resulted from feeding the amoebae on specific seaweeds. Cuticles of live Sargassum and Macrocystis were digested easily by the amoebae. However cuticles of autoclaved Macrocystis and those of Porphyra (fresh or autoclaved) were eaten last. In spite of the absence of identifiable extracellular enzymatic activity in the medium the amoebae were able to cut and consume live fronds and blocks of gelled agars carrageenans, and alginates.     

External barium affects the gating of KCNQ1 potassium channels and produces a pore block via two discrete sites

The pore properties and the reciprocal interactions between permeant ions and the gating of KCNQ channels are poorly understood. Here we used external barium to investigate the permeation characteristics of homomeric KCNQ1 channels. We assessed the Ba(2+) binding kinetics and the concentration and voltage dependence of Ba(2+) steady-state block. Our results indicate that extracellular Ba(2+) exerts a series of complex effects, including a voltage-dependent pore blockade as well as unique gating alterations. External barium interacts with the permeation pathway of KCNQ1 at two discrete and nonsequential sites. (a) A slow deep Ba(2+) site that occludes the channel pore and could be simulated by a model of voltage-dependent block. (b) A fast superficial Ba(2+) site that barely contributes to channel block and mostly affects channel gating by shifting rightward the voltage dependence of activation, slowing activation, speeding up deactivation kinetics, and inhibiting channel inactivation. A model of voltage-dependent block cannot predict the complex impact of Ba(2+) on channel gating in low external K(+) solutions. Ba(2+) binding to this superficial site likely modifies the gating transitions states of KCNQ1. Both sites appear to reside in the permeation pathway as high external K(+) attenuates Ba(2+) inhibition of channel conductance and abolishes its impact on channel gating. Our data suggest that despite the high degree of homology of the pore region among the various K(+) channels, KCNQ1 channels display significant structural and functional uniqueness.     

STUDIES OF VANDADIUM-BROMOPEROXIDASE USING SURFACE AND CORTICAL PROTOPLASTS OF MACROCYSTIS PYRIEFERA (PHAEOPHYTA)1

Aqueous Tri-SO₄ buffer (pH 8.3) extracts of cortical and surface protoplasts of Macrocystis pyrifera (L.) C. Ag. Catalyzed the bromination of monochlorodimedone (2-chloro-5, 5-dimethyl-1, 3-dimedone, MCD) in the presence of hydrogen peroxide and bromide. The apparent bromo-peroxidase activity as measured by the bromination of MCD was inhibited by the presence of endogenous compounds which are probably polyphenolics compounds (i.e. polymers of phloroglucinol) or other inhibitors. The bromoperoxidase activity of the protoplast extracts increased substantially when the extracts were washed extensively with Tris-SO₄ buffer (pH 8.3) by ultrafiltration. The bromoperoxidase activity of both surface and cortical protoplast extracts was dependent on the presence of vanadium, indicating that the bromoperoxidase present in cortical and surface cells of M. pyrifera is vanadium-bromoperoxidase. Halogenated compounds constitute one of the most significant classes of marine natural products. Since bromoperoxidases are assumed to be involved in the biosynthesis of these compounds, elucidation of the location of BrPO with in the algal tissue is important.     

An inactivation gate in the selectivity filter of KCNQ1 potassium channels

Inactivation is an inherent property of most voltage-gated K⁺ channels. While fast N-type inactivation has been analyzed in biophysical and structural details, the mechanisms underlying slow inactivation are yet poorly understood. Here, we characterized a slow inactivation mechanism in various KCNQ1 pore mutants, including L273F, which hinders entry of external Ba²⁺ to its deep site in the pore and traps it by slowing its egress. Kinetic studies, molecular modeling, and dynamics simulations suggest that this slow inactivation involves conformational changes that converge to the outer carbonyl ring of the selectivity filter, where the backbone becomes less flexible. This mechanism involves acceleration of inactivation kinetics and enhancement of Ba²⁺ trapping at elevated external K⁺ concentrations. Hence, KCNQ1 slow inactivation considerably differs from C-type inactivation where vacation of K⁺ from the filter was invoked. We suggest that trapping of K⁺ at s₁ due to filter rigidity and hindrance of the dehydration-resolvation transition underlie the slow inactivation of KCNQ1 pore mutants.     

Biotransformation of alkanes and haloalkanes by a marine amoeba

Summary A marine amoeba, Trichosphaerium I-7, originally found feeding on macroalgae in a region of natural oil seepage, was maintained in the laboratory for prolonged periods on hexadecane, octadecane, 1-chlorooctadecane, or 1-bromooctadecane as a carbon source. The cells attached readily and eroded holes in thin layers of these compounds. Crystalline and clear spherical inclusions appeared in the cytoplasm of cells fed these xenobiotics followed by a marked cell darkening. Thin layer chromatographic analyses of acetone extracts from amoebae grown for 12 days on [1-¹⁴C]octadecane demonstrated the formation of labelled substances of higher polarity than the original alkane. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography revealed that ¹⁴C derived from [1-¹⁴C]octadecane was incorporated into acetone-insoluble macromolecules.Offprint requests to: D. D. Kaska     

Tunicamycin-sensitive glycoproteins involved in the mating of Chlamydomonas reinhardi

It has been previously shown that mild trypsinization of Chlamydomonas gametes reversibly inhibits steps of the mating process. Gametic agglutination is delayed 30–60 min, while cell wall hydrolysis and zygote formation are delayed 1–3 h. If gametes are pretreated with 5 μg/ml tunicamycin (TM) for 1 h and then trypsinized, the recovery of agglutination is blocked. These results indicate that N-glycosylated glycoproteins are involved in agglutination. Treatment of normal gametes with tunicamycin alone does not have a significant effect on agglutination and mating efficiency, suggesting that there is little or no turnover of the surface receptors before mating. Tunicamycin also interferes with cell growth and prevents the conversion of vegetative cells into gametes.