Discovery and Characterization of Peptide Inhibitors for Calcium and Integrin Binding Protein 1

Autor(es):Mostrar menos -

Puhl, Ana C. ^[1] ; Bogart, Jonathan W. ^[1] ; Haberman, Victoria A. ^[1] ; Larson, Jacob E. ^[1] ; Godoy, Andre S. ^[2] ; Norris-Drouin, Jacqueline L. ^[1] ; Cholensky, Stephanie H. ^[1] ; Leisner, Tina M. ^[3] ; Frye, V, Stephen ; Parise, V, Leslie ; Bowers, Albert A. ^{[4, 5, 6]} ; Pearce, Kenneth H. ^{[5, 6]}

Afiliação do(s) autor(es)

^[1] Univ N Carolina, Ctr Integrat Chem Biol & Drug Discovery, Eshelman Sch Pharm, Chem Biol & Med Chem, Chapel Hill, NC 27599 - USA

^[2] Univ Sao Paulo, Inst Phys Sao Carlos, BR-13563120 Sao Carlos - Brazil

^[3] V, Univ N Carolina, Dept Biochem & Biophys, Chapel Hill, NC 27599 - USA

^[4] Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 - USA

^[5] Frye, Stephen, V, Univ N Carolina, Ctr Integrat Chem Biol & Drug Discovery, Eshelman Sch Pharm, Chem Biol & Med Chem, Chapel Hill, NC 27599 - USA

^[6] Parise, Leslie, V, Frye, Stephen, V, UNC Lineberger Comprehens Canc Ctr, Chapel Hill, NC 27599 - USA

• Tipo de documento: Artigo Científico
• Fonte: ACS Chemical Biology; v. 15, n. 6, p. 1505-1516, JUN 19 2020.
• Citações Web of Science: 0

Resumo

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 CIBI with low nanomolar affinity. CIB1 was cocrystallized with UNC10245092, and the 2.1 angstrom resolution structure revealed that the peptide binds as an alpha-helix in the H10 pocket, displacing the CIBI 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. (AU)

• Processo FAPESP: 16/19712-9 - Caracterização estrutural das proteínas do vírus Zika e busca por agentes antivirais
• Beneficiário: Andre Schutzer de Godoy
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado