| Autor: |
Singh SA; Department of Pathology, Children's Hospital Boston and Harvard Medical School, Boston, Massachusetts, USA., Winter D, Bilimoria PM, Bonni A, Steen H, Steen JA |
| Jazyk: |
angličtina |
| Zdroj: |
Nature methods [Nat Methods] 2012 Apr 08; Vol. 9 (5), pp. 504-8. Date of Electronic Publication: 2012 Apr 08. |
| DOI: |
10.1038/nmeth.1970 |
| Abstrakt: |
We introduce a mass spectrometry-based method that provides residue-resolved quantitative information about protein phosphorylation. In this assay we combined our full-length expressed stable isotope-labeled protein for quantification strategy (FLEXIQuant) with a traditional kinase assay to determine the mechanisms of multikinase substrate phosphorylation such as priming-dependent kinase activities. The assay monitors the decrease in signal intensity of the substrate peptides and the concomitant increase in the (n × 80 Da)-shifted phosphorylated peptide. We analyzed the c-Jun N-terminal kinase (JNK)-dependent glycogen synthase kinase 3β (GSK3β) activity on doublecortin (DCX) revealing mechanistic details about the role of phosphorylation cross-talk in GSK3β activity and permitting an advanced model for GSK3β-mediated signaling. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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