Determination of Binding Potency of Peptidic Inhibitors of Grb2-SH2 by Using the Protein-Captured Biosensor Method
| Autor: | Hui-Ming Chen, Wan-Ching Li, Feng-Di T. Lung, Yung-Hsien Chang |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Bioanalysis
Recombinant Fusion Proteins Peptide SH2 domain Biochemistry Antibodies Antigen-Antibody Reactions Structural Biology Amino Acid Sequence Surface plasmon resonance GRB2 Adaptor Protein Glutathione Transferase chemistry.chemical_classification biology Chemistry General Medicine Surface Plasmon Resonance Combinatorial chemistry Protein Structure Tertiary Costs and Cost Analysis biology.protein GRB2 biological phenomena cell phenomena and immunity Signal transduction Oligopeptides Biosensor Protein Binding Proto-oncogene tyrosine-protein kinase Src |
| Zdroj: | Protein & Peptide Letters. 15:806-810 |
| ISSN: | 0929-8665 |
| DOI: | 10.2174/092986608785203683 |
| Popis: | The growth factor receptor-binding protein 2 – Src homology 2 (Grb2 – SH2) domain plays an important role in the oncogenic Ras signal transduction pathway, therefore, peptidic inhibitors of the Grb2 – SH2 domain has been chosen as our target for the development of antiproliferative agents. The inhibitory effects of peptide analogs on the Grb2 – SH2 domain have been determined by using surface plasmon resonance (SPR) technology developed with the BIACORE biosensor. Recently, we reported the analysis of interactions between peptides and the GST-Grb2-SH2 that was immobilized on the surface of sensor chip by using BIACORE biosensor (the protein-immobilized method). Herein, we analyze interactions of peptides with the GST-Grb2-SH2 that was captured by the anti-GST antibodies immobilized on the surface of sensor chip (the protein-captured method). Results obtained by both methods are in good correlation, indicating the immobilization of GST-Grb2-SH2 on the sensor chip did not significantly affect the binding of Grb2-SH2 with peptides. Both SPR-based assays are very sensitive bioanalytical methods and can be applied in screening inhibitors of target proteins or purifying GST-fusion proteins, however, considering the efficiency and the cost, the GST-Grb2-SH2-immobilized method is suggested for routinely determining the binding potency of inhibitors of Grb2-SH2. |
| Databáze: | OpenAIRE |
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