Structure of tubulin C‐terminal domain obtained by subtilisin treatment The major α and β tubulin isotypes from pig brain are glutamylated

FEBS Letters

Redeker, Virginie²  Rossier, Jean²  Melki, Ronald¹  Le Caer, Jean-Pierre²  Promé, Danielle³ 

[1]Laboratoire d'Enzymologie, Centre National de la Recherche Scientifique, 91198 Gif-sur-Yvette Cedex, France

[2]Institut Alfred Fessard, Centre National de la Recherche Scientifique, 91198 Gif-sur-Yvette Cedex, France

[3]Centre de Recherche de Biochimie et Génétique Cellulaires, Centre National de la Recherche Scientifique, 31062 Toulouse Cedex, France

关键词: Glutamylation;    Mass spectrometry;    Post-translational modification;    Subtilisin;    Tubulin;    FAB-MS;    fast atom bombardment mass spectrometry;    HPLC;    high-performance liquid chromatography;    IEF;    isoelectric focusing;    PMSF;    phenylmethylsulfonyl fluoride;    SDS-PAGE;    sodium dodecyl sulfate polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(92)81441-N

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Limited subtilisin digestion of the tubulin α, β heterodimer has been used in this work to reduce the total number of tubulin isotypes from 20 for native to 9 for subtilisin-cleaved tubulin. This indicates that the major part of tubulin heterogeneity is located at the C-terminus of the molecule. The C-terminal peptides of both α and β subunits of tubulin were purified by anion-exchange HPLC. Combined use of Edman degradation chemistry and mass spectrometry on the isolated peptides shows that subtilisin cleavage occurs at position Asp-438 and His-406 of α and Gln-433 and His-396 of β tubulin chains. Quantitative analysis of our data show that cleavage at positions His-406 (α) and His-396 (β) occurs with a low efficiency and indicates that the major isotypes of pig brain tubulin are modified by sequential attachment of 1 to 5 glutamic acid residues at positions Glu-445 or −435 of α and β tubulin, respectively.

【 授权许可】

Unknown