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Related Concept Videos

Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

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...
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

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...
Phosphorylation01:02

Phosphorylation

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...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

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 the...

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Updated: May 29, 2026

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
11:23

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

Detecting protein higher-order structural changes using kinase as a phospho-labeler.

Asato Maeda1, Kosuke Ogata1, Yasushi Ishihama2

  • 1Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan.

Cell Reports Methods
|May 27, 2026
PubMed
Summary
This summary is machine-generated.

This study introduces a novel method for detecting protein structural changes using phosphate labeling and quantitative phosphoproteomics. This technique offers site-specific insights into protein dynamics across the entire proteome.

Keywords:
CP: biotechnologyin vitro kinase reactionmass spectrometryphosphopeptide enrichmentphosphoproteomicsphosphorylationprotein structurestructural proteomics

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Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate
11:31

Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate

Published on: September 18, 2013

Related Experiment Videos

Last Updated: May 29, 2026

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
11:23

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate
11:31

Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate

Published on: September 18, 2013

Area of Science:

  • Biochemistry
  • Proteomics
  • Structural Biology

Background:

  • Understanding protein structure and dynamics is crucial for cellular function.
  • Existing methods for assessing proteome-wide structural changes are limited.
  • Phosphorylation plays a key role in regulating protein structure and function.

Purpose of the Study:

  • To develop a residue-specific method for detecting proteome-wide protein structural alterations.
  • To profile structural dynamics within the intracellular proteome.
  • To correlate phosphorylation efficiency with substrate protein structure.

Main Methods:

  • Developed a structural proteomics approach combining in vitro kinase reactions with residue-specific labeling.
  • Employed quantitative phosphoproteomics with phosphopeptide enrichment for site-resolved profiling.
  • Utilized myoglobin and HEK293T cell extracts under various conditions (heat denaturation, protease treatment, RNA digestion).

Main Results:

  • Demonstrated that phosphorylation efficiency accurately reflects differences in substrate protein structure.
  • Successfully identified proteome-wide structural changes in non-denatured cell extracts by comparing phosphorylation efficiencies before and after RNA digestion.
  • Validated the approach using myoglobin and cell extracts, showing sensitivity to structural variations.

Conclusions:

  • The developed approach enables a residue-specific readout of structural dynamics within the intracellular proteome.
  • This method provides a powerful tool for investigating protein structural changes on a proteome-wide scale.
  • The findings highlight the link between phosphorylation and protein structure, offering new avenues for research.