Posttranslational Modifications in the C-terminal Tail of Axonemal Tubulin from Sea Urchin Sperm (*)
- Jean Mary(1)(§),
- Virginie Redeker(1),
- Jean-Pierre Le Caer(1),
- Jean Rossier(1) and
- Jean-Marie Schmitter(2)
- From the (1)Laboratoire de Neurobiologie de la Diversité Cellulaire, CNRS URA 2054, ESPCI, 10 rue Vauquelin, 75231 Paris cedex 5, France and the
- (2)Laboratoire de Biochimie, CNRS URA 1970, Ecole Polytechnique, 91128 Palaiseau cedex, France
- § This work forms a portion of the doctoral thesis of J. Mary, who received a predoctoral fellowship from the Ministère de l'Education Nationale et de l'Enseignement Supérieur et de la Recherche (MENESR, France). To whom correspondence should be addressed. Tel.: 33-1-40-79-47-65; Fax: 33-1-40-79-47-57; jean.mary{at}espci.fr.
Abstract
After proteolytic digestion of sperm tubulin from sea urchin Paracentrotus lividus, C-terminal peptides were isolated by chromatographic separations. The peptides were analyzed by Edman degradation and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. About 70% of the isolated C-terminal peptides were unmodified. The remaining modified peptides have undergone a combination of numerous posttranslational modifications generating significant heterogeneity of sperm tubulin. α-Tubulin is modified by detyrosylation, release of the penultimate glutamate, polyglutamylation, and polyglycylation. Glycylation and glutamylation can coexist within one α-tubulin isoform. β-Tubulin undergoes polyglycylation but was not observed to be polyglutamylated. The number of units posttranslationally added reaches 11 and 12 glycyl units on β- and α-tubulin, respectively. This is different from the polyglycylation of axonemal tubulin in Paramecium cilia where up to 40 added glycyl units were observed both on α- and β-tubulin.
Footnotes
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↵* This work was supported by the Association pour la Recherche contre le Cancer (ARC, France) and the Institut National de la Santé et de la Recherche Médicale (INSERM CRE/930808). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵1 J. Mary, C. Klotz, V. Redeker, J.-P. Le Caer, J. Rossier, and J.-M. Schmitter, unpublished results.
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↵2 The abbreviations used are:
- HPLC
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high performance liquid chromatography
- DEAE
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diethylaminoethyl
- MALDI-TOF
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matrix-assisted laser desorption/ionization-time of flight
- Pipes
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1,4-piperazine diethanesulfonic acid.
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- Received January 22, 1996.
