How does studied phosphorylation?
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Protein phosphorylation is studied through various methods and approaches. One approach involves the purification and manipulation of phosphoproteins, using computational and biochemical techniques to analyze their structure and function . Another strategy combines fluorescence resonance energy transfer (FRET) with a small molecule ATP analogue to study protein phosphorylation in living cells . Phosphorylation can also be examined through PAGE electrophoresis and tandem mass spectrometry (MS/MS) to identify specific phosphorylation sites . Additionally, a novel method based on the phosphorylation-dependent electrophoretic mobility shift (PDEMS) has been developed to detect protein phosphorylation, allowing for the measurement of the ratio of protein phosphorylation in cells . These different approaches provide insights into the mechanisms and functional implications of protein phosphorylation in various cellular processes .
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| The paper describes the process of studying protein phosphorylation, including demonstrating phosphorylation, identifying phosphorylation sites, determining the functional implications of phosphorylation, and using phosphorylation sites in vivo. | |
29 Citations | The paper describes a method to detect protein phosphorylation using a phosphorylation-dependent electrophoretic mobility shift (PDEMS) in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The PDEMS is caused by the distribution of negatively charged amino acids around the phosphorylation site, resulting in a shift in protein mobility during SDS-PAGE. |
Open access•Journal Article | The paper discusses the methods of analyzing protein phosphorylation, including examining changes in protein mobility during PAGE electrophoresis and using tandem mass spectrometry to identify specific phosphorylation sites. |
3 Citations | The paper studied protein phosphorylation by combining fluorescence resonance energy transfer (FRET) with a small molecule adenosine 5′-triphosphate (ATP) analogue. They used a new ATP analogue called ATP-NB to phosphorylate proteins in solution and in living cells, and then labeled them with a tetrazine modified fluorescent probe called TZ-Cy3. |
25 Citations | The paper proposes a general workflow for protein phosphorylation analysis that combines computational and biochemical approaches. It highlights the use of Molecular Dynamics simulations as a decision-making tool to design the most appropriate phosphomimetic variant. |
Related Questions
What’s the importance of phosphorylation in the cell?5 answersProtein phosphorylation plays a crucial role in cellular function and signaling. It serves as a key post-translational modification that regulates protein activity, impacting signal transduction, gene expression, cell cycle progression, and various cellular processes. Dysregulation of phosphorylation is linked to the development of diseases, particularly cancer, making kinase inhibitors valuable for therapeutic interventions. Phosphorylation creates distinct protein phosphoforms with diverse biological consequences, necessitating the study of defined phosphoforms to understand their functions. Specificity in phosphorylation is achieved through the spatial organization of kinases and phosphatases within cells, forming signaling complexes that regulate cellular responses. Overall, phosphorylation is a critical mechanism that influences protein function, cellular behavior, and disease pathogenesis, highlighting its significance in cell biology.
What is phosphorelay signal transduction system?4 answersPhosphorelay signal transduction systems are mechanisms that allow microorganisms to sense and respond to environmental changes through the modification of gene expression. These systems involve the transfer of a phosphate group between specific histidine and aspartic residues in proteins. They are found in various organisms, including bacteria and eukaryotes, and play a crucial role in regulating important cellular processes such as stress response, virulence, and biofilm development. The phosphorelay systems can consist of multiple components, including sensor proteins, intermediate proteins, and response regulators, which work together to transmit and amplify signals. Understanding the function and regulation of phosphorelay systems is important for gaining insights into the evolution and adaptation of organisms to their environments.
Why dephosphorylate a plasmid?3 answersDephosphorylating a plasmid is done to prevent recircularization of the plasmid during ligation procedures. This is important because recircularization can interfere with subsequent steps in molecular biology and genetic engineering, such as gene expression and transfection. By removing the 5'-phosphate residues from the termini of the plasmid, the recircularization process is suppressed, allowing for more efficient ligation and preventing unwanted recombination events. Additionally, dephosphorylation of the plasmid can enhance stability and transfection efficiency, as it prevents polyubiquitination and degradation of the plasmid. Overall, dephosphorylation of a plasmid is a crucial step in molecular biology and genetic engineering to ensure successful downstream applications and maximize the efficiency of gene expression and transfection.
Can phosphorylation inhibit apoptosis induction?3 answersPhosphorylation can both inhibit and induce apoptosis depending on the context. In the case of nucleolin, phosphorylation at specific sites (Thr-76 and 84) was found to be involved in protecting against myocardial ischemia/reperfusion (I/R) injury. On the other hand, hexokinase-dependent retrotranslocation of truncated BID was shown to be a selective protective mechanism against death receptor-induced apoptosis on the mitochondria. Ponicidin was found to induce apoptosis in leukemia cells by downregulating survivin and Bcl-2 expressions. Additionally, oTR was shown to induce apoptosis in SMMC-7721 cells through the activation of intrinsic mitochondria-mediated pathways. These findings suggest that phosphorylation can have both pro-apoptotic and anti-apoptotic effects depending on the specific protein and cellular context.
What is a protein kinase and how is protein phosphorylation used during signal transduction?5 answersProtein kinases are enzymes that add phosphate groups to other proteins through a process called phosphorylation. This modification can lead to functional changes in the target protein, such as altering enzyme activity, cellular location, or association with other proteins. Protein kinases play a crucial role in signal transduction, which is the process by which cells communicate and respond to external stimuli. They regulate the majority of cellular pathways, including those involved in intracellular signaling transduction cascades. Protein phosphorylation is also important in the induction and maintenance of pluripotency in stem cells, as well as in the generation of induced pluripotent stem cells (iPSCs). In addition, protein kinases contribute to signal transduction during the induction of ischemia tolerance and damage following cerebral ischemia, with specific kinases like protein kinase A, protein kinase B (Akt), and protein kinase C playing important roles. Furthermore, protein kinases are involved in the regulation of circadian rhythms, including the entrainment of the master circadian clock to light-dark transitions. Overall, protein kinases are essential for cellular signaling and the regulation of various biological processes.
What does ERK phosphorylate?2 answersERK phosphorylates a variety of proteins involved in different cellular processes. It has been shown to phosphorylate extracellular signal-regulated kinase (ERK) itself, as well as the cell survival protein 90-kDa ribosomal S6 kinase (RSK)1. ERK also phosphorylates transcription factors and chromatin modifying enzymes, thereby regulating gene expression. Additionally, ERK phosphorylation is involved in the regulation of cell proliferation, cell survival, cell growth, cell metabolism, cell migration, and cell differentiation. The phosphorylation of ERK is crucial for conveying growth and mitogenic signals, and dysregulation of ERK phosphorylation has been implicated in various human diseases. ERK phosphorylation is also associated with drug addiction, as chronic drug and alcohol exposure can modify ERK phosphorylation within central reinforcement systems.