Analyzing protein tyrosine phosphatases by phosphotyrosine analog integration.
Reversible protein phosphorylation plays a central role in cellular signal transduction and is a focus of biomedical studies. However, it is a challenging task to study the effects of protein phosphorylation in the presence of protein phosphatase activities, especially for protein tyrosine phosphatases SHP1, SHP2 and LMW-PTP, which are themselves regulated by protein tyrosine phosphorylation. Expressed protein ligation, by combining chemical peptide synthesis with recombinant protein expression, allows for site-specific unnatural modifications of semisynthetic proteins. In this review, we describe how semisynthetic proteins were prepared to incorporate nonhydrolyzable phosphotyrosine analogs, and utilized in combination with site-directed mutagenesis and other means to elucidate regulatory mechanisms of protein tyrosine phosphatases.
Shen
Kui
K
Department of Chemistry and Biochemistry, Northern Illinois University, DeKalb, IL 60115, USA. kshen@niu.edu
D000595:Amino Acid Sequence; D051380:GRB2 Adaptor Protein / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019000:Phosphotyrosine / Q000031:analogs & derivatives* / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D025941:Protein Interaction Mapping / Q000379:methods; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6 / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000032:analysis / Q000378:metabolism*; D015398:Signal Transduction
D051380:GRB2 Adaptor Protein; D047908:Intracellular Signaling Peptides and Proteins; D019000:Phosphotyrosine; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases