23 research outputs found
Unbiased Functional Proteomics Strategy for Protein Kinase Inhibitor Validation and Identification of bona fide Protein Kinase Substrates: Application to Identification of EEF1D as a Substrate for CK2
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Differential Proteomic Analysis of Mammalian Tissues Using SILAM
Differential expression of proteins between tissues underlies organ-specific functions. Under certain pathological conditions, this may also lead to tissue vulnerability. Furthermore, post-translational modifications exist between different cell types and pathological conditions. We employed SILAM (Stable Isotope Labeling in Mammals) combined with mass spectrometry to quantify the proteome between mammalian tissues. Using 15N labeled rat tissue, we quantified 3742 phosphorylated peptides in nuclear extracts from liver and brain tissue. Analysis of the phosphorylation sites revealed tissue specific kinase motifs. Although these tissues are quite different in their composition and function, more than 500 protein identifications were common to both tissues. Specifically, we identified an up-regulation in the brain of the phosphoprotein, ZFHX1B, in which a genetic deletion causes the neurological disorder MowatβWilson syndrome. Finally, pathway analysis revealed distinct nuclear pathways enriched in each tissue. Our findings provide a valuable resource as a starting point for further understanding of tissue specific gene regulation and demonstrate SILAM as a useful strategy for the differential proteomic analysis of mammalian tissues
Two Distinct Mechanisms for Actin Capping Protein RegulationβSteric and Allosteric Inhibition
A crystallographic study reveals the structural basis for regulation by two different inhibitors of the actin capping protein, a critical factor controlling actin-driven cell motility
Mixing Standard Work and Nonstandard Deals: The Consequences of Heterogeneity in Employment Arrangements
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Headspace vapor characterization of Hanford waste tank 241-U-108: Results from samples collected on 8/29/95
This report describes the analytical results of vapor samples taken from the headspace of the waste storage tank 241-U-108 (Tank U-108) at the Hanford Site in Washington State. The results described in the report were obtained to characterize the vapors present in the tank headspace and to support safety evaluations and tank farm operations. The results include air concentrations of selected inorganic and organic analytes and grouped compounds from samples obtained by Westinghouse Hanford Company (WHC) and provided for analysis to Pacific Northwest National Laboratory (PNNL). Analyte concentrations were based on analytical results and, where appropriate, sample volumes provided by WHC
Serine 23 and 36 Phosphorylation of Caveolin-2 Is Differentially Regulated by Targeting to Lipid Raft/Caveolae and in Mitotic Endothelial Cells
Protein kinase CK2: Systematic relationships with other posttranslational modifications
Β© Springer International Publishing Switzerland 2015. A wealth of biochemical and genetic evidence has demonstrated that protein kinase CK2 has critical roles in the regulation and execution of numerous biological processes. Large-scale proteomic and phosphoproteomics studies have further reinforced the widespread impact of CK2 on cellular events through interactions with many cellular proteins or protein complexes and through phosphorylation of a vast number of cellular proteins. Given its global participation in many fundamental processes, it is not surprising that CK2 has been implicated in numerous human diseases, a factor that has spurred interest in CK2 as a candidate for molecular- targeted therapy. Despite this growing profile, many questions regarding its precise mechanisms of regulation remain. In fact, several lines of evidence suggest that CK2 is constitutively active, leading to a speculation that CK2 is an unregulated enzyme. Accordingly, there is an apparent paradox that leads to the question of how an unregulated enzyme such as CK2 can be a participant in regulatory processes. In an effort to resolve this paradox, studies in our lab and others have focused on an investigation of the relationships between CK2 and other cellular pathways. Using a combination of computational predictions and database mining together with proteomic strategies and biochemical assays, we have been elucidating systematic relationships between CK2 and regulatory pathways where CK2 phosphorylation sites overlap other posttranslational modifications. Overall, these studies suggest intriguing mechanisms by which CK2 can participate in regulatory events and also how alterations in CK2 levels that accompany disease may promote pathological rewiring of regulatory pathways