Effects of FTDP‐17 mutations on the in vitro phosphorylation of tau by glycogen synthase kinase 3β identified by mass spectrometry demonstrate certain mutations exert long‐range conformational changes

FEBS Letters

Gallo, Jean-Marc²  Gibb, Graham M²  Hutton, Michael¹  Betts, Joanna C³  Blackstock, Walter P³  Connell, James W²  Anderton, Brian H²  Lovestone, Simon² 

[1] The Birdsall Research Building, Mayo Clinic Jacksonville, 4500 San Pablo Road, Jacksonville, FL 32224, USA;Department of Neuroscience, Institute of Psychiatry, King's College London, De Crespigny Park, London SE5 8AF, UK;Biomolecular Structure Unit, GlaxoSmithKline, Research and Development, Stevenage SG1 2NY, UK

关键词: Tau;    Phosphorylation;    Frontotemporal dementia with Parkinsonism linked to chromosome 17;    Glycogen synthase kinase 3β;    Two dimensional phosphopeptide mapping;    Mass spectrometry;    FTDP-17;    frontotemporal dementia with Parkinsonism linked to chromosome 17;    GSK3β;    glycogen synthase kinase 3β;    PHF;    paired helical filament;    nanoES-MS;    nanoelectrospray mass spectrometry;    DTT;    dithiothreitol;    PMSF;    phenylmethylsulphonylfluoride;    SDS–PAGE;    sodium dodecyl sulphate–polyacrylamide electrophoresis;   

DOI  :  10.1016/S0014-5793(01)02267-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In vitro phosphorylation of recombinant wild-type 2N4R tau and FTDP-17 exonic mutant forms P301L, V337M and R406W by glycogen synthase kinase 3β (GSK3β) was examined by two dimensional phosphopeptide mapping analysis on thin layer cellulose plates. Comparison of these peptide maps with those generated from wild-type 1N4R tau isoform from which the phosphopeptide constituents and sites of phosphorylation had been determined previously, enabled us to monitor directly changes in phosphorylation of the individual tau proteins. No differences were found in the phosphorylation of wild-type, P301L or V337M tau by GSK3β but the R406W mutant showed at least two clear differences from the other three tau proteins. The peptides, identified by mass spectrometry corresponding to phosphorylation at both threonine 231 and serine 235 (spot 3), serines 396, 400 and 404 (spot 6a) and serines 195 and 199 (spot 6b) were absent from the R406W peptide map. The findings imply that the R406W mutation in tau exerts long-range conformational effects on the structure of tau.

【 授权许可】

Unknown