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L V Evans

Publications and source records attributed to L V Evans.

10 recordsLinked to original sources

Mucilaginous substances from macroalgae: an overview.

Macroalgae (seaweeds) produce a diversity of mucilaginous substances, some of which are of considerable commercial and biotechnological importance (e.g. alginate, carrageenan, agar). They are involved in vivo in conferring structural integrity as cell wall materials (alginate, agar, carrageenan), the prevention of desiccation (sulphated fucans), selective absorption of ions (alginate, sulphated fucans) and in bioadhesion (various glycoproteins and mucopolysaccharides). The current state of knowledge with respect to the structure, localization, biosynthesis and secretion of some of the more important of these substances is summarised.

Carbohydrate Sequence↗

Fertilization in brown algae. V. Further investigations of lectins as surface probes.

The binding of FITC- (fluorescein isothiocyanate), Au- and 125I-labelled lectins (conA, RCA120 (Ricinus communis agglutinin, MW 120 000) and FBP (fucose-binding protein from Lotus tetragonobolus)) to gametes of Fucus serratus and their physiological effects on fertilization have been studied. Results indicate that eggs strongly bind FITC- and Au-labelled conA and RCA120, whilst FITC-FBP binds strongly to sperm. All three iodinated lectins bound to eggs but this was apparently non-specific and similar in magnitude to the binding of iodinated bovine serum albumin. The results suggested the possibility of two distinct types of lectin receptor on egg surfaces: non-specific, highly abundant receptors and less abundant, specific receptors, possibly locally aggregated. All three lectins inhibit fertilization, FBP being the most effective.

Eukaryota↗

Fertilization in brown algae. IV. Appearance of sperm-specific antigens on fertilized eggs.

Flagella antigens from sperm of Fucus serratus have been used to raise antibodies in rabbits. The immunoglobulin G fraction inhibits fertilization with some degree of species specificity. The antigens detected on sperm are not present on unfertilized egg membranes, but appear after fertilization. The common antigens on the fertilized egg can be distinguished from the cell wall material that is also released on fertilization.

Epitopes↗

Fertilization in brown algae. III. Preliminary characterization of putative gamete receptors from eggs and sperm of Fucus serratus.

Membrane fractions have been isolated from eggs of Fucus serrratus which inhibit fertilization in a species-specific manner. This activity is destroyed by alpha-fucosidase and alpha-mannosidase. Some 6% of the protein of this membrane fraction binds to Con A-agarose, following SDS solubilization, and when eluted with alpha-methyl mannoside inhibits fertilization when preincubated with sperm, but not eggs. This inhibitory activity is species-specific and destroyed by alpha-fucosidase but not by trypsin. SDS-gel electrophoresis reveals 1 band staining strongly with Coomassie Brilliant Blue G and weakly with the PAS reagent. This major band represents a glycoprotein with an approximate molecular weight of 30 000 Daltons. Membrane fractions from sperm of Fucus serratus solubilized in KC1 yielded a protein-containing fraction, after affinity chromatography on desulphated focoidan-Sepharose. This fraction is 100-fold more effective in inhibiting fertilization after preincubation with eggs than either Con A or fucose-binding protein. It is species-specific and inhibition is reversed when pretreated eggs are washed with fucoidan. Activity is destroyed by heat and trypsin and only one diffuse band is apparent on SDS gels. This stains positively with Coomassie Brilliant Blue G but not with PAS and has a molecular weight of approximately 60 000 Daltons. Tentative calculation of the numbers of putative receptor molecules gives a figure of 2.5 X 10(9) receptors per egg and 1.8 X 10(6) receptors per sperm.

Cell Membrane↗

Fertilization in brown algae. II. Evidence for lectin-sensitive complementary receptors involved in gamete recognition in Fucus serratus.

Fertilization in Fucus serratus is directly proportional to the number of sperm added, saturating at approximately 250 sperm per egg with an apparent Km of 120 sperm per egg. The effect of a range of lectins on fertilization has been tested. Preincubation of gametes with Con A and fucose-binding protein (FBP) inhibited fertilization. At low concentrations this was by specifically binding to eggs; at high concentrations pretreatment of either gametes inhibited fertilization probably due to cytotoxicity. Fertilization was not inhibited by simple sugar haptens, but polysaccharides containing fucosyl or mannosyl residues (yeast mannan, fucoidan, ascophyllan) inhibited fertilization by binding to sperm. Pretreatment of eggs with alpha-fucosidase or alpha-mannosidase was effective in inhibiting fertilization. All the results indirectly demonstrate that fertilization in Fucus serratus is based on an association between fucosyl- and mannosyl-containing ligands on the egg surface and specific carbohydrate-binding receptors on the sperm surface.

Carbohydrates↗

The role of Golgi bodies in polysaccharide sulphation in Fucuszygotes.

The cell wall of 24-h zygotes of Fucus serratus is composed of 3 layers--an inner fibrillar layer (sulphated fucan), an outer fibrillar layer (alginic aicd/cellulose) and an exterior amorphous layer (sulphated fucan, alginic acid). The 2 layers containing sulphated fucan are preferentially thickened at the rhizoid pole. Light- and electron-microscope autoradiographic pulse-chase experiments on 22-h zygotes using 35SO2-(4) show the Golgi bodies to be the sites of fucan sulphation. The isolation and characterization of isolated Golgi-rich fractions from 22-h zygotes shows that the first detectable labelled macromolecule is associated with these fractions 2 min after addition of 35SO2-(4). The sulphate acceptor molecule has been partially characterized. 35S-APS and 35S-paps are detectable in the soluble fraction 0.5 min after addition of 35SO2-(4). The results are discussed in relation to other published work on the differentiation of Fucus embryos and on polysaccharide sulphation.

Autoradiography↗

Fertilization in brown algae. I. SEM and other observations on Fucus serratus.

The cell wall secreted immediately following sperm entry into an egg can be visualized by the fluorescent dye Calcofluor white. Cell wall secretion precedes nuclear fusion by 10-20 min. SEM observations of the surface of unfertilized and fertilized eggs and sperm attachment to eggs are described. These results are discussed in relation to fertilization in sea urchins and the biochemical phenomena associated with egg-sperm recognition in Fucus.

Cell Wall↗