TY - JOUR
T1 - Characterization of posttranslational modifications in neuron-specific class III β-tubulin by mass spectrometry
AU - Alexander, Janice E.
AU - Hunt, Donald F.
AU - Lee, Michael K.
AU - Shabanowitz, Jeffrey
AU - Michel, Hanspeter
AU - Berlin, Sunetary C.
AU - Macdonald, Timothy L.
AU - Sundberg, Richard J.
AU - Rebhun, Lionel I.
AU - Frankfurter, Anthony
PY - 1991/6/1
Y1 - 1991/6/1
N2 - Class III β-tubulin, isolated from adult bovine brain, is resolved into at least seven charge variants on isoelectric focusing gels. To identify the posttranslational modifications responsible for this heterogeneity, a mixture of brain tubulins was treated with cyanogen bromide and the C-terminal fragments from the class III β-tubulin isoforms were then isolated by binding them to the monoclonal antibody TuJ1. Combined use of tandem mass spectrometry and both subtractive and automated Edman degradation chemistry on the isolated peptides indicates that many of the isoforms differ by phosphorylation at Ser-444 plus attachment of one to six glutamic acid molecules to the side chain of the first glutamate residue, Glu-438, in the C-terminal sequence Tyr-Glu-Asp-Asp-Glu-Glu-Glu-Ser-Glu-Ala-Gln-Gly-Pro-Lys.
AB - Class III β-tubulin, isolated from adult bovine brain, is resolved into at least seven charge variants on isoelectric focusing gels. To identify the posttranslational modifications responsible for this heterogeneity, a mixture of brain tubulins was treated with cyanogen bromide and the C-terminal fragments from the class III β-tubulin isoforms were then isolated by binding them to the monoclonal antibody TuJ1. Combined use of tandem mass spectrometry and both subtractive and automated Edman degradation chemistry on the isolated peptides indicates that many of the isoforms differ by phosphorylation at Ser-444 plus attachment of one to six glutamic acid molecules to the side chain of the first glutamate residue, Glu-438, in the C-terminal sequence Tyr-Glu-Asp-Asp-Glu-Glu-Glu-Ser-Glu-Ala-Gln-Gly-Pro-Lys.
KW - Isoelectric focusing
KW - Tandem mass spectrometry
KW - Tubulin heterogeneity
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U2 - 10.1073/pnas.88.11.4685
DO - 10.1073/pnas.88.11.4685
M3 - Article
C2 - 2052551
AN - SCOPUS:0025845285
SN - 0027-8424
VL - 88
SP - 4685
EP - 4689
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 11
ER -