Thermal stabilization of tissues and the preservation of protein phosphorylation states for two-dimensional gel electrophoresis
| Autor: | Alexander J. Trachtenberg, Jae-Hyung Robert Chang, Winston Patrick Kuo, Gary B. Smejkal, Emily A. Freeman, Alexander R. Ivanov, Alexander Lazarev, Chiara Rivas-Morello |
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| Rok vydání: | 2010 |
| Předmět: |
Clinical Biochemistry
Phosphatase Molecular Sequence Data Biology Biochemistry Analytical Chemistry Dephosphorylation Mice Tandem Mass Spectrometry Catalytic Domain Image Processing Computer-Assisted Animals Protein phosphorylation Electrophoresis Gel Two-Dimensional Trypsin Amino Acid Sequence Kinase activity Phosphorylation chemistry.chemical_classification Cerebral Cortex Analysis of Variance Two-dimensional gel electrophoresis Kinase Protein Stability Protein-Tyrosine Kinases Phosphoproteins Peptide Fragments Mice Inbred C57BL Enzyme chemistry Chromatography Liquid |
| Zdroj: | Electrophoresis. 32(16) |
| ISSN: | 1522-2683 |
| Popis: | 2-DE is typically capable of discriminating proteins differing by a single phosphorylation or dephosphorylation event. However, a reliable representation of protein phosphorylation states as they occur in vivo requires that both phosphatases and kinases are rapidly and completely inactivated. Thermal stabilization of mouse cerebral cortex homogenates effectively inactivated these enzymes, as evidenced by comparison with unstabilized tissues where abscissal pI shifts were a common feature in 2-D gels. Of the 588 matched proteins separated on 2-D gels comparing stabilized and unstabilized tissues, 53 proteins exhibited greater than twofold differences in spot volume (ANOVA, p |
| Databáze: | OpenAIRE |
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