Determination of the endogenous phosphorylation state of B‐50/GAP‐43 and neurogranin in different brain regions by electrospray mass spectrometry

FEBS Letters

Caputi, Antonio³  Cattabeni, Flaminio³  Gispen, Willem H.²  Di Luca, Monica³  Pastorino, Lucia³  Raverdino, Vittorio¹  De Graan, Pierre N.E.² 

[1] EMC-Analytical Laboratory, Hewlett Packard, 39 rue de Veyrot, 1217 Meyrin, Switzerland;Rudolf Magnus Institute for Neurosciences, University of Utrecht, Universiteitsweg 100, 3508 TA Utrecht, The Netherlands;Institute of Pharmacological Sciences, University of Milano, via Balzaretti 9, 20133 Milano, Italy

关键词: Synaptic plasticity;    Protein kinase C;    Hippocampus;    Cortex;    Phosphoprotein;    (Rat);    ES/MS;    electrospray mass spectrometry;    HPLC;    high-pressure liquid chromatography;    LTP;    long-term potentiation;    PCA;    perchloric acid;    PKC;    protein kinase C;    SDS-PAGE;    sodium dodecyl sulphate-polyacrylamide gel electrophoresis;    TFA;    trifluoroacetic acid;   

DOI  :  10.1016/0014-5793(96)00612-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Electrospray mass spectrometry coupled to liquid chromatography was utilized to measure two PKC neuronal substrates, B-50/GAP-43 and neurogranin, in single rat brain areas. Aliquots of perchloric acid extracts were directly injected and mass spectra recorded. At elution times of 14.2 and 27.0 min two molecular species of MW 7450 and 23 602 Da were observed. These values are in excellent agreement for the expected MW for rat neurogranin and B-50/GAP-43. The presence of molecular species shifted by 80 mass units in both cases indicates that these proteins are present in phosphorylated forms in cortical and hippocampal extracts.

【 授权许可】

Unknown