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Hitting the target: emerging technologies in the search for kinase substrates
Brendan D Manning1, Lewis C Cantley
1Department of Cell Biology, Harvard Medical School, Division of Signal Transduction, Beth Israel Deaconess Medical Center, 4 Blackfan Circle, Boston, MA 02115, USA.
Science'S STKE : Signal Transduction Knowledge Environment
|December 12, 2002
Summary
Identifying protein kinase substrates is crucial for cell biology. New methods like phage display and phospho-motif antibodies offer more reliable ways to find genuine in vivo kinase substrates, overcoming limitations of traditional techniques.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Protein kinases regulate cellular functions through phosphorylation.
- Identifying specific protein kinase substrates is essential but challenging.
- Traditional methods for substrate identification are often laborious and unreliable.
Purpose of the Study:
- To review and discuss novel techniques for identifying protein kinase substrates.
- To highlight the utility and limitations of emerging substrate identification strategies.
Main Methods:
- Screening phage expression libraries for phosphoproteins.
- Peptide library screens to identify kinase-specific phosphorylation motifs.
- Utilizing phospho-motif antibodies.
- Employing engineered kinases with ATP analogs and inhibitors.
Main Results:
- New approaches have demonstrated success in identifying genuine in vivo kinase substrates.
- These methods offer improved reliability and efficiency compared to traditional techniques.
- Each method has unique advantages and inherent caveats that require consideration.
Conclusions:
- Novel methodologies are advancing the field of kinase substrate identification.
- These advanced techniques are critical for a comprehensive understanding of kinase signaling pathways.
- Further development and application of these methods will accelerate discoveries in cell signaling.