Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Assaying protein kinase activity.

Jan Brábek1, Steven K Hanks

  • 1Department of Cell and Developmental Biology, Vanderbuilt University School of Medicine, Nashville, TN, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 3, 2004
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Integrated transcriptomic and proteomic analysis of cancer-suppressive Mesocestoides corti larvae.

Parasites & vectors·2026
Same author

Novel soluble bazedoxifene formulation gains antiproliferative and migrastatic effect in squamous cell carcinoma cells.

Scientific reports·2026
Same author

Cancer invasion plasticity during the metastatic cascade: strategies to survive and thrive.

Cancer letters·2026
Same author

Design and Synthesis of Actin-Targeting 10-Phenoxy Cytochalasan Analogues: Balancing Cytotoxicity and Migrastatic Activity.

ACS medicinal chemistry letters·2026
Same author

Migrastatic therapy as a potential game-changer in adaptive cancer treatment.

Scientific reports·2026
Same author

The potential of cytochalasins and other cytochalasans as drugs targeting cancer cell invasiveness and metastasis.

European journal of medicinal chemistry·2025
Same journal

Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

High-Performance Computing in Tandem Mass Spectrometry (MS/MS) Peptide Identification.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Engineering and Adapting Disulfide-Containing Proteins to Enable Intracellular Functionality.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

AI-Driven Protein Research: From Prediction to Design.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

This study details methods for measuring protein kinase activity, focusing on the oncogenic protein-tyrosine kinase v-Src. Protocols cover expression, purification, and activity assays, crucial for understanding cell signaling pathways.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Signaling

Background:

  • Protein kinases regulate crucial cellular functions and are encoded by approximately 2% of eukaryotic genes.
  • Assessing protein kinase catalytic activity is vital for elucidating signal-transduction pathways.
  • The oncogenic protein-tyrosine kinase v-Src serves as a model for studying kinase activity.

Purpose of the Study:

  • To present methods for determining the activity of the protein-tyrosine kinase v-Src.
  • To provide protocols for heterologous expression, purification, and kinase assays.
  • To offer guidance on assaying other protein kinases, considering substrate specificity and buffer conditions.

Main Methods:

  • Heterologous expression of v-Src in yeast (Saccharomyces cerevisiae).

Related Experiment Videos

  • Immunoaffinity purification of v-Src from yeast cell lysates.
  • Kinase reactions involving 32P incorporation into peptide substrates and activity quantification.

    • Main Results:

    • Established protocols for the expression and purification of active v-Src in yeast.
    • Demonstrated methods for quantifying protein kinase activity using radiolabeled substrates.
    • Provided a framework for adapting these methods to other protein kinases.

    Conclusions:

    • The presented methods enable robust assessment of protein kinase activity, exemplified by v-Src.
    • Accurate measurement of kinase activity is essential for dissecting complex cell signaling networks.
    • Recommendations are provided for optimizing kinase assays based on enzyme and substrate characteristics.