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Related Concept Videos

Protein Networks02:26

Protein Networks

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An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
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Interactions Between Signaling Pathways01:19

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Signaling cascades usually lack linearity. Multiple pathways interact and regulate one another, allowing cells to integrate and respond to diverse environmental stimuli.
Convergence and divergence, and cross-talk between signaling pathways
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Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
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Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...
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Related Experiment Video

Updated: Aug 11, 2025

Quantification of Protein Interaction Network Dynamics using Multiplexed Co-Immunoprecipitation
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Quantification of Protein Interaction Network Dynamics using Multiplexed Co-Immunoprecipitation

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Multiplexed kinase interactome profiling quantifies cellular network activity and plasticity.

Martin Golkowski1, Andrea Lius1, Tanmay Sapre1

  • 1Department of Pharmacology, University of Washington, Seattle, WA 98195, USA.

Molecular Cell
|February 3, 2023
PubMed
Summary
This summary is machine-generated.

We developed a new method, kiCCA, to map protein-protein interactions (PPIs) for kinases. This approach reveals how kinase networks change in cancer, identifying potential drug targets like AAK1 for cancer plasticity and drug resistance.

Keywords:
kinomenative complexprotein interactionsproteomicssignaling pathways

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Identification of Kinase-substrate Pairs Using High Throughput Screening
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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Proteomics

Background:

  • Protein-protein interaction (PPI) networks are crucial for cellular functions and disease pathogenesis.
  • Kinases are key regulatory components within these networks, but studying their dynamic interactions remains challenging.
  • Existing methods lack the speed and multiplexing capability for systematic analysis of kinase interactome dynamics.

Purpose of the Study:

  • To develop and validate a novel chemoproteomic method for profiling kinase interactome dynamics.
  • To investigate context-dependent changes in kinase interactomes across diverse cancer types.
  • To identify novel kinase-associated drug targets and understand their role in cancer progression.

Main Methods:

  • Kinobead competition and correlation analysis (kiCCA), a quantitative mass spectrometry-based chemoproteomic technique.
  • High-throughput, multiplexed profiling of endogenous kinase interactomes.
  • Analysis of kinase PPIs across 18 cancer cell lines to assess context-specific alterations.

Main Results:

  • Identification of 1,154 protein-protein interactions involving 238 kinases.
  • Quantification of dynamic kinase interactome changes correlated with cancer type, plasticity, and signaling states.
  • Discovery of novel drug target candidates, including the endocytic adapter-associated kinase 1 (AAK1) complex.

Conclusions:

  • The kiCCA method enables rapid and comprehensive profiling of kinase interactome dynamics.
  • Kinase interactome plasticity is a significant factor in cancer cell signaling, progression, and drug resistance.
  • Targeting specific kinase complexes, such as AAK1, offers potential therapeutic strategies for cancer treatment.