Creation of Phosphotyrosine Superbinders by Directed Evolution of an SH2 Domain.

Posted on January 15, 2017

Title	Creation of Phosphotyrosine Superbinders by Directed Evolution of an SH2 Domain.
Publication Type	Journal Article
Year of Publication	2017
Authors	Huang, Haiming, Kaneko Tomonori, Sidhu Sachdev S., and Li Shawn S. C.
Journal	Methods Mol Biol
Volume	1555
Pagination	225-254
Date Published	2017
ISSN	1940-6029
Keywords	Amino Acid Sequence, Binding Sites, Carrier Proteins, Cell Surface Display Techniques, Computational Biology, Conserved Sequence, Enzyme-Linked Immunosorbent Assay, Evolution, Molecular, Genetic Vectors, Kinesis, Models, Molecular, Mutagenesis, Site-Directed, Peptide Library, Phosphorylation, Phosphotyrosine, Position-Specific Scoring Matrices, Protein Binding, Protein Conformation, Protein Interaction Domains and Motifs, Sequence Analysis, DNA, Software, src Homology Domains, Workflow
Abstract	<p>Commercial antibodies raised against phosphotyrosine have been widely used as reagents to detect or isolate tyrosine-phosphorylated proteins from cellular samples. However, these antibodies are costly and are not amenable to in-house production in an academic lab setting. In this chapter, we describe a method to generate super-high affinity SH2 domains, dubbed the phosphotyrosine superbinders, by evolving a natural SH2 domain using the phage display technology. The superbinders are stable and can be easily produced in Escherichia coli in large quantities. The strategy presented here may also be applied to other protein domains to generate domain variants with markedly enhanced affinities for a specific post-translational modification.</p>
DOI	10.1007/978-1-4939-6762-9_13
Alternate Journal	Methods Mol. Biol.
PubMed ID	28092036