TTLL10 can perform tubulin glycylation when co-expressed with TTLL8.
Tubulin can undergo unusual post-translational modifications, glycylation and glutamylation. We previously failed to find glycylase (glycine ligase) for tubulin while identifying TTLL10 as a polyglycylase for nucleosome assembly protein 1. We here examine whether TTLL10 performs tubulin glycylation. We used a polyclonal antibody (R-polygly) raised against a poly(glycine) chain, which does not recognize monoglycylated protein. R-polygly strongly reacted with mouse tracheal cilia and axonemal tubulins. R-polygly detected many proteins in cell lysates co-expressing TTLL10 with TTLL8. Two-dimensional electrophoresis revealed that the R-polygly-reactive proteins included alpha- and beta-tubulin. R-polygly labeling signals overlapped with microtubules. These results indicate that TTLL10 can strongly glycylate tubulin in a TTLL8-dependent manner. Furthermore, these two TTLL proteins can glycylate unidentified 170-, 110-, 75-, 40-, 35-, and 30-kDa acidic proteins.
Department of Molecular Anatomy, Molecular Imaging Advanced Research Center, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan.
Koji
K
Ikegami
Mitsutoshi
M
Setou
D000818:Animals; D000918:Antibody Specificity; D054468:Axoneme / Q000378:metabolism; D019556:COS Cells; D002522:Chlorocebus aethiops; D002923:Cilia / Q000378:metabolism; D015870:Gene Expression; D006031:Glycosylation; D007150:Immunohistochemistry; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D010453:Peptide Synthases / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014132:Trachea / Q000378:metabolism; D014404:Tubulin / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*
D011994:Recombinant Proteins; D014404:Tubulin; D010453:Peptide Synthases; C112698:tubulin polyglutamylase