Surface organization and composition of Euglena. II. Flagellar mastigonemes

Abstract

The surface of the Euglena flagellum is coated with about 30,000 fine filaments of two distinct types. The longer of these nontubular mastigonemes (about 3 micron) appear to be attached to the paraflagellar rod whereas the shorter nontubular mastigonemes (about 1.5 micron) are the centrifugally arranged portions of a larger complex, which consists of an attached unit parallel to and outside of the flagellar membrane. Units are arranged laternally in near registration and longitudinally overlap by one-half of a unit length. Rows of mastigoneme units are firmly attached to the axoneme microtubules or to the paraflagellar rod as evidenced by their persistence after removal of the flagellar membrane with neutral detergents. SDS-acrylamide gels of whole flagella revealed about 30 polypeptides, of which two gave strong positive staining with the periodic acid-Schiff (PAS) procedure. At least one of these two bands (glycoproteins) has been equated with the surface mastigonemes by parallel analysis of isolated and purified mastigonemes, particularly after phenol extraction. The faster moving glycoprotein has been selectively removed from whole flagella and from the mastigoneme fraction with low concentrations of neutral detergents at neutral or high pH. The larger glycoprotein was found to be polydisperse when electrophoresed through 1% agarose/SDS gels. Thin-layer chromatography of hydrolysates of whole flagella or of isolated mastigonemes has indicated that the major carbohydrate moiety is the pentose sugar, xylose, with possibly a small amount of glucose and an unknown minor component.

Scale-associated glycoproteins of Scherffelia dubia (Chlorophyta) form high-molecular-weight complexes between the scale layers and the flagellar membrane

“…Some examples are; the large cell-body scales of prymnesiophyte algae (recently reviewed by Leadbeater 1994), the flagellar (Becker et al 1990;Becker and Melkonian 1992) and thecal scales (Becker et al 1989(Becker et al , 1995c of Tetraselmis striata, and the basket scales of Mesostigma viride (Domozych et al 1991). In addition, different types of flagellar hairs have been biochemically characterized: from Chlamydomonas reinhardii (Witman et al 1972;Monk et al 1983), Euglena gracilis (Bouck et al 1978;Rogalski and Bouck 1980) and Ochromonas (Kawano and Bouck 1984). In most of these studies only a few glycoproteins were found and, so far, the presence of disulfide bridges has not been investigated.…”

Characterization and localization of a flagellar-specific membrane glycoprotein in Euglena.

“…Previous studies (3) have revealed that treatment of whole flagella with low concentrations of detergents selectively extracts one of the two major glycoproteins and leaves the other major glycoprotein(s) (mastigonemes) in a particulate fraction . The following method therefore provided soluble extracts of the one flagellar glycoprotein with minimal disruption of the axoneme, membrane, and mastigonemes.…”

Fluorescent mannosides serve as acceptor substrates for glycosyltransferase and sugar-1-phosphate transferase activities in Euglena gracilis membranes

“…Dark grown culture after seven days was harvested by centrifugation (800 g for 5 min), washed twice with deionised water and once in HEPES buffer (10 mM, pH 7.0 HEPES/KOH, 25 mM KCl). The harvested cells were re-suspended in HEPES buffer and de-flagellated by the method of cold-shock [30] that required incubation on ice for 2 h and the collection of de-flagellated cells by centrifugation (800 g for 5 min).…”