Abstract
Extraction of demembranated bull sperm flagella by SDS was used to maximize tubulin solubilization. The α- and β-tubulin separated by SDS-PAGE were treated with endoproteinases LysC and AspN, respectively. Carboxy-terminal fragments were isolated by Mono Q chromatography and reversed-phase HPLC. Automated sequencing and mass spectrometry revealed an astonishingly high number of tubulin variants. Many variants were due to polyglutamylation and in particular to polyglycylation. The number of side-chain glycyl residues ranged from 0 to 28 in α and 0 to 15 in β. Corresponding values for side-chain glutamyl residues were 0–6 in α and 0–3 in β. Additional α variability was based on carboxy-terminal detyrosination and partial loss of the penultimate glutamate. A major glycylation site in α- and β-tubulin was mapped. Some variants seem to display both glycyl and glutamyl side chains.
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Plessmann, U., Weber, K. Mammalian Sperm Tubulin: An Exceptionally Large Number of Variants Based on Several Posttranslational Modifications. J Protein Chem 16, 385–390 (1997). https://doi.org/10.1023/A:1026332621215
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DOI: https://doi.org/10.1023/A:1026332621215