Purification and characterization of basal apparatuses from a flagellate green alga

Distribution of polyglutamylated tubulin in the flagellar apparatus of green flagellates

“…In axonemes, detyrosinated tubulin is highly enriched in the B-tubules constituting of about two thirds of the total ␣-tubulin of this subfiber Johnson, 1998]. Thus, the contamination would have to be tremendous, which was not confirmed by EM analysis of our preparations [Geimer et al, 1997]. Further, using L3 we generally reproduced the results described by Johnson [1998] using a different antibody to detyrosinated tubulin [SG, Gunderson et al, 1984] (e.g., low amounts of detyrosinated tubulin in the flagellar tip region of Fig.…”

“…Accordingly, L3 could partially cross-react with polyglutamylated tubulin, which would be prevented if these epitopes were blocked by GT335. The weak staining of the basal bodies with L3 is even more puzzling because our previous biochemical analysis has shown that 30% of the ␣-tubulin in basal apparatus preparations was detyrosinated or ␦2-tubulin [Geimer et al, 1997]. We offer two potential explanations for this contradiction: (1) the detyrosinated tubulin in the basal bodies cannot be accessed or recognized by L3, or (2) the preparations of basal apparatus used for the biochemical analysis were contaminated with detyrosinated tubulin (e.g., from remnant axonemal microtubules).…”

“…e: L3 epitopes mostly blocked as in the sheet of doublets in Figure 5c. polyglutamylated ␣-tubulin with short side chains were detected in axonemes [Geimer et al, 1997]. Polyglycylation is characteristic for axonemal tubulin but absent in the transition region or basal bodies [Pechart et al, 1999;Million et al, 1999].…”

“…Often basal bodies or centrioles are enriched in glutamylated tubulin [Fouquet et al, 1994;DelgadoViscogliosi et al, 1996;Bobinnec et al, 1998a]. In the green alga Spermatozopsis similis, polyglutamylated ␣-tubulin with side chains of up to 17 residues is enriched in preparations of basal bodies [Geimer et al, 1997].…”

Localization of Golgi 58K protein (formiminotransferase cyclodeaminase) to the centrosome

“…Gold particles were seen near the appendages, but were not concentrated in these structures in this study. About 5% of the a-tubulin carries a polyglutamyl side chain that can reach at least 17 residues in length in basal bodies from the naked green flagellate Spermatozopsis similes (Geimer et al 1997). In the adult mammalian brain, more than 90% of a-tubulin is glutamylated, containing Fig.…”