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Expression and Production of SH2 Domain Proteins
Bernard A Liu1, Mari Ogiue-Ikeda2, Kazuya Machida3
1Broad Institute of MIT and Harvard, 415 Main St., 5175 JJ, Cambridge, MA, 02142, USA.
Methods in Molecular Biology (Clifton, N.J.)
|January 17, 2017
Summary
This study details methods for producing soluble recombinant Src Homology 2 (SH2) domain proteins. These proteins are essential tools for studying phosphotyrosine signaling pathways and protein interactions.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Signaling
Background:
- Src Homology 2 (SH2) domains are crucial for phosphotyrosine signaling.
- SH2 domains mediate interactions downstream of receptor tyrosine kinases (RTKs).
- Mammals possess over a hundred SH2 domains with distinct specificities.
Purpose of the Study:
- To describe methods for producing soluble recombinant SH2 domain proteins.
- To provide protocols for SH2 domain purification for various biochemical studies.
- To facilitate research into the role of SH2 domains in phosphotyrosine signaling.
Main Methods:
- Expression and purification of SH2 domains using GST or poly His-tags.
- Stepwise protocols for recombinant protein production.
- Validation studies for assessing protein quality.
Main Results:
- Established methods for producing purified SH2 domain proteins.
- Detailed protocols applicable to a wide range of SH2 domains.
- Characterization of purified human SH2 domains.
Conclusions:
- Soluble recombinant SH2 proteins are essential tools for phosphotyrosine signaling research.
- The described methods enable the production of high-quality SH2 domain proteins.
- These proteins support diverse proteomic and biochemical applications.
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