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Receptor Tyrosine Kinases

Receptor Tyrosine Kinases

Receptor tyrosine kinases or RTKs are membrane-bound receptors that phosphorylate specific tyrosine on protein substrates. RTKs regulate cellular growth, differentiation, survival, and migration. They contain an extracellular ligand binding domain, a transmembrane domain, and a cytosolic tail with intrinsic kinase activity. Several extracellular signaling molecules activate RTKs in one or more ways and relay the signal downstream. Ligands such as platelet-derived growth factor (PDGF) or...

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Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

Published on: February 21, 2019

Protocol to characterize extracellular c-Src tyrosine kinase function through substrate interaction and

Michael P Omini¹, Deema Alsalih², SarahBeth D Votra²

¹Department of Urology, SUNY Upstate Medical University, Syracuse, NY 13210, USA; Department of Biology, College of Arts and Science at Syracuse University, Syracuse, NY 13244, USA.

|December 3, 2023

Summary

This study introduces a new protocol to identify extracellular tyrosine-phosphorylated substrates secreted by cancer cells. This method aids in understanding the role of extracellular Src (e-Src) in cancer progression.

Keywords:

Cancer Cell Biology Molecular Biology

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Last Updated: Jul 9, 2025

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

Published on: February 21, 2019

Identification of Kinase-substrate Pairs Using High Throughput Screening

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

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

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

Published on: June 30, 2019

Area of Science:

Biochemistry
Cell Biology
Oncology

Background:

Cellular Src tyrosine kinase (c-Src) is found in the secretomes of human cancers, termed extracellular Src (e-Src).
Phosphoproteomics identified 114 potential extracellular e-Src substrates, including Tissue Inhibitor of Metalloproteinases 2.

Purpose of the Study:

To present a detailed protocol for characterizing secreted tyrosine-phosphorylated substrates resulting from c-Src expression and secretion.
To provide a method adaptable for studying other extracellular kinases.

Main Methods:

Collection of cell secretomes and extracts.
Antibody treatment and Ni-NTA pull-down assays.
Detection of protein-protein interactions and tyrosine phosphorylation of substrates.

Main Results:

A comprehensive protocol is described for identifying extracellular substrates of c-Src.
The method allows for the characterization of tyrosine-phosphorylated proteins in the extracellular environment.
The protocol facilitates the study of e-Src's role in cancer secretomes.

Conclusions:

The developed protocol enables the characterization of secreted tyrosine-phosphorylated substrates linked to c-Src.
This methodology is valuable for investigating the function of extracellular kinases in various biological contexts, particularly in cancer research.
The protocol offers a foundation for future studies on extracellular kinase signaling pathways.