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Related Concept Videos

Enzymes02:34

Enzymes

Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...
Introduction to Mechanisms of Enzyme Catalysis01:13

Introduction to Mechanisms of Enzyme Catalysis

For many years, scientists thought that enzyme-substrate binding took place in a simple "lock-and-key" fashion. This model stated that the enzyme and substrate fit together perfectly in one instantaneous step. However, current research supports a more refined view scientists call induced fit. The induced-fit model expands upon the lock-and-key model by describing a more dynamic interaction between enzyme and substrate. As the enzyme and substrate come together, their interaction causes a mild...
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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Related Experiment Video

Updated: Jun 27, 2026

Assaying Protein Kinase Activity with Radiolabeled ATP
08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

Tuning a three-component reaction for trapping kinase substrate complexes.

Alexander V Statsuk1, Dustin J Maly, Markus A Seeliger

  • 1Howard Hughes Medical Institute, Department of Cellular and Molecular Pharmacology, University of California, San Francisco, California 94107, USA.

Journal of the American Chemical Society
|December 5, 2008
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel chemical cross-linking method to identify unknown protein kinases. This technique enables selective targeting of kinases in complex cellular environments, advancing phosphoproteome research.

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Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
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Identification of Kinase-substrate Pairs Using High Throughput Screening

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Related Experiment Videos

Last Updated: Jun 27, 2026

Assaying Protein Kinase Activity with Radiolabeled ATP
08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

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

Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

Area of Science:

  • Biochemistry
  • Chemical Biology
  • Proteomics

Background:

  • Identifying upstream protein kinases is crucial for understanding phosphorylation events in the phosphoproteome.
  • Existing chemical methods for studying kinase-substrate interactions face challenges with specificity in complex biological samples.

Purpose of the Study:

  • To develop a selective chemical cross-linking strategy for identifying protein kinases and their substrates within cell lysates.
  • To overcome limitations of non-specific reactions encountered with earlier cross-linking agents.

Main Methods:

  • A three-component chemical reaction was designed to covalently link substrate-kinase complexes.
  • The cross-linker was modified by replacing the adenosine moiety with a kinase inhibitor scaffold and the o-phthaldialdehyde with a thiophene-2,3-dicarboxaldehyde.
  • The modified cross-linker was tested for its ability to selectively target protein kinases in the presence of competing cellular proteins.

Main Results:

  • The initial cross-linker (1) showed non-specific side reactions in complex cellular environments.
  • Structural modifications to the cross-linker significantly improved its selectivity for protein kinases.
  • The optimized cross-linker successfully achieved covalent cross-linking between a cysteine-containing substrate and its corresponding kinase in cell lysates.

Conclusions:

  • The modified chemical cross-linker provides a highly selective method for identifying protein kinases.
  • This approach facilitates the discovery of previously unknown upstream kinases involved in phosphorylation.
  • The study advances techniques for phosphoproteome analysis and kinase-substrate mapping.