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Related Concept Videos

Phosphorylation01:02

Phosphorylation

49.9K
The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
49.9K
Regulation of Nuclear Protein Sorting01:45

Regulation of Nuclear Protein Sorting

2.4K
Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
2.4K
Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

6.8K
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
6.8K
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

13.0K
Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
13.0K
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

8.3K
When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze...
8.3K
Disassembly of Intermediate Filaments01:35

Disassembly of Intermediate Filaments

2.0K
Intermediate filaments (IFs) do not undergo spontaneous disassembly. Enzymes, kinases, and phosphatases add and remove phosphates from specific sites to regulate their disassembly. The IF concentration in the cytoplasm also regulates the disassembly. If the concentration crosses a threshold, it activates the protein kinases in the vicinity, allowing the phosphorylation of IFs.
Keratin proteins, found at the cell periphery near cell junctions, undergo a cycle of assembly and disassembly. In Type...
2.0K

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Related Experiment Video

Updated: Jun 6, 2025

Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis
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Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis

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Phosphorylation-mediated conformational change regulates human SLFN11.

Michael Kugler1, Felix J Metzner1, Gregor Witte1

  • 1Gene Center and Department of Biochemistry, Ludwig-Maximilians-Universität München, Feodor-Lynen Straße 25, 81377, Munich, Germany.

Nature Communications
|December 3, 2024
PubMed
Summary

Human Schlafen 11 (SLFN11) protein regulates DNA damage response and viral defense. New cryo-EM structures reveal how phosphorylation controls SLFN11

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Assessing Cellular Target Engagement by SHP2 PTPN11 Phosphatase Inhibitors
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Related Experiment Videos

Last Updated: Jun 6, 2025

Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis
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Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis

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Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
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Assessing Cellular Target Engagement by SHP2 PTPN11 Phosphatase Inhibitors
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Area of Science:

  • Molecular Biology
  • Structural Biology
  • Biochemistry

Background:

  • Human Schlafen 11 (SLFN11) is a protein that sensitizes cells to DNA damaging agents, acting as a predictive biomarker in chemotherapy.
  • SLFN11 also functions as an antiviral restriction factor by recognizing single-stranded DNA (ssDNA) and inhibiting translation.
  • The precise regulation of SLFN11's diverse functions and enzymatic activities has remained unclear.

Purpose of the Study:

  • To elucidate the structural mechanisms underlying SLFN11 function and regulation.
  • To investigate the role of phosphorylation in modulating SLFN11's interaction with tRNAs and ssDNA.
  • To understand how SLFN11's nuclease activity is controlled.

Main Methods:

  • Cryo-electron microscopy (cryo-EM) was used to determine the structures of SLFN11 bound to specific tRNAs.
  • Site-directed mutagenesis was employed to create phosphomimetic mutants (e.g., S753D).
  • Biochemical assays were performed to assess ssDNA binding, ATP binding, and ribonuclease activity.

Main Results:

  • Cryo-EM structures reveal how SLFN11 binds and cleaves tRNA-Leu and tRNA-Met, with regulation by phosphorylation at S219 and T230.
  • The S753D phosphomimetic mutant exhibits a monomeric conformation, binds ATP, but loses ssDNA binding and shows reduced ribonuclease activity.
  • Phosphorylation at S753 acts as a conformational switch, governing SLFN11 dimerization and binding of ATP and ssDNA.

Conclusions:

  • Phosphorylation at S753 is critical for regulating SLFN11 dimerization and substrate binding (ATP and ssDNA).
  • Phosphorylation at S219 and T230 specifically modulates tRNA recognition and nuclease activity.
  • These findings provide a structural basis for understanding the multifaceted roles of SLFN11 in DNA damage response and antiviral immunity.