title: Protein methylation in full length Chlamydomonas flagella.
abstract: Post-translational protein modification occurs extensively in eukaryotic flagella. Here we examine protein methylation, a protein modification that has only recently been reported to occur in flagella [Schneider MJ, Ulland M, Sloboda RD.2008. Mol Biol Cell 19(10):4319-4327.]. The cobalamin (vitamin B12) independent form of the enzyme methionine synthase (MetE), which catalyzes the final step in methionine production, is localized to flagella. Here we demonstrate, using immunogold scanning electron microscopy, that MetE is bound to the outer doublets of the flagellum. Methionine can be converted to S-adenosyl methionine, which then serves as the methyl donor for protein methylation reactions. Using antibodies that recognize symmetrically or asymmetrically methylated arginine residues, we identify three highly methylated proteins in intact flagella: two symmetrically methylated proteins of about 30 and 40 kDa, and one asymmetrically methylated protein of about 75 kDa. Several other relatively less methylated proteins could also be detected. Fractionation and immunoblot analysis shows that these proteins are components of the flagellar axoneme. Immunogold thin section electron microscopy indicates that the symmetrically methylated proteins are located in the central region of the axoneme, perhaps as components of the central pair complex and the radial spokes, while the asymmetrically methylated proteins are associated with the outer doublets. Cell Motil. Cytoskeleton 2009. (c) 2009 Wiley-Liss, Inc.
authors.affiliation: Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire, USA. rds@dartmouth.edu
authors.forename: Roger D
authors.initials: RD
authors.lastname: Sloboda
authors.forename: Louisa
authors.initials: L
authors.lastname: Howard
mesh_terms: D013766:5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase / Q000378:metabolism / Q000648:ultrastructure; D000818:Animals; D002696:Chlamydomonas / Q000378:metabolism* / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D005407:Flagella / Q000378:metabolism* / Q000648:ultrastructure; D015151:Immunoblotting; D007150:Immunohistochemistry; D008745:Methylation; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D008954:Models, Biological; D015800:Protozoan Proteins / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D015800:Protozoan Proteins; D013766:5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase