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Structure of tubulin C‐terminal domain obtained by subtilisin treatment The major α and β tubulin isotypes from pig brain are glutamylated
Author(s) -
Redeker Virginie,
Melki Ronald,
Promé Danielle,
Le Caer Jean-Pierre,
Rossier Jean
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)81441-n
Subject(s) - tubulin , subtilisin , chemistry , terminal (telecommunication) , domain (mathematical analysis) , c terminus , biochemistry , microbiology and biotechnology , biophysics , biology , microtubule , amino acid , enzyme , computer science , mathematics , telecommunications , mathematical analysis
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.