Discovery and characterization of peptide inhibitors for calcium and integrin binding protein 1
| Autor: | Andre S. Godoy, Victoria A Haberman, Ana C. Puhl, Leslie V. Parise, Jacob E. Larson, Stephen V. Frye, Jonathan W. Bogart, Kenneth H. Pearce, Tina M. Leisner, Jacqueline Norris-Drouin, Stephanie H. Cholensky, Albert A. Bowers |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Phage display Protein Conformation MEMBRANA PLASMÁTICA Peptide Calorimetry 01 natural sciences Biochemistry Article 03 medical and health sciences Protein structure Cell Line Tumor Drug Discovery Humans Amino Acid Sequence Peptide sequence Integrin binding chemistry.chemical_classification 010405 organic chemistry Chemistry Calcium-Binding Proteins Isothermal titration calorimetry General Medicine 0104 chemical sciences 030104 developmental biology Förster resonance energy transfer Cell-penetrating peptide Molecular Medicine Hydrophobic and Hydrophilic Interactions |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP ACS Chem Biol |
| Popis: | Calcium and integrin binding protein 1 (CIB1) is an EF-hand-containing, small intracellular protein that has recently been implicated in cancer cell survival and proliferation. In particular, CIB1 depletion significantly impairs tumor growth in triple-negative breast cancer (TNBC). Thus, CIB1 is a potentially attractive target for cancer chemotherapy that has yet to be validated by a chemical probe. To produce a probe molecule to the CIB1 helix 10 (H10) pocket and demonstrate that it is a viable target for molecular intervention, we employed random peptide phage display to screen and select CIB1-binding peptides. The top peptide sequence selected, UNC10245092, was produced synthetically and binding to CIB1 was confirmed by isothermal titration calorimetry (ITC) and a time-resolved fluorescence resonance energy transfer (TR-FRET) assay. Both assays showed that the peptide bound to CIB1 with low nanomolar affinity. CIB1 was co-crystallized with UNC10245092, and the 2.1 Å resolution structure revealed that the peptide binds as an α-helix in the H10 pocket, displacing the CIB1 C-terminal H10 helix and causing conformational changes in H7 and H8. UNC10245092 was further derivatized with a C-terminal Tat-derived cell penetrating peptide (CPP) to demonstrate its effects on TNBC cells in culture, which are consistent with results of CIB1 depletion. These studies provide a first-in-class chemical tool for CIB1 inhibition in cell culture and validate the CIB1 H10 pocket for future probe and drug discovery efforts. |
| Databáze: | OpenAIRE |
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