Posttranslational modifications of axonemal tubulin.
Axonemal tubulin exhibits a high degree of heterogeneity mostly due to several posttranslational modifications (PTM). The aim of this work was to chemically characterize the different PTM occurring in the C-terminal tail of axonemal tubulin purified from sea urchin, Paracentrotus lividus, spermatozoa. After its purification, tubulin was enzymatically cleaved. The C-terminal peptides were chromatographically isolated, first by anion exchange and then by reverse-phase HPLC. Peptides were characterized by their sequence, determined by Edman degradation, and by their mass, determined by MALDI-TOF/MS. The two major conclusions are that the majority of the isolated C-terminal peptides were unmodified and that polyglycylation and polyglutamylation can occur simultaneously on one molecule of alpha-tubulin.
Mary
J
J
Laboratoire de Neurobiologie et Diversité Cellulaire, CNRS URA 2054, ESPCI, Paris, France. jean.mary@espci.fr
Redeker
V
V
Le Caer
J P
JP
Rossier
J
J
Schmitter
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D007525:Isoelectric Focusing; D008297:Male; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis / Q000302:isolation & purification; D011499:Protein Processing, Post-Translational; D012617:Sea Urchins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013082:Sperm Tail / Q000737:chemistry / Q000378:metabolism*; D013094:Spermatozoa / Q000378:metabolism; D014404:Tubulin / Q000302:isolation & purification / Q000378:metabolism*
D010446:Peptide Fragments; D014404:Tubulin