Epi/pericellular protein disulfide isomerase: a novel regulator of biomechanical a...
| Grant number: | 14/23657-8 |
| Support type: | Scholarships abroad - Research Internship - Post-doctor |
| Effective date (Start): | March 15, 2015 |
| Effective date (End): | March 14, 2016 |
| Field of knowledge: | Health Sciences - Medicine |
| Principal Investigator: | Francisco Rafael Martins Laurindo |
| Grantee: | Ana Iochabel Soares Moretti |
| Supervisor abroad: | Hervé Kovacic |
| Home Institution: | Instituto do Coração Professor Euryclides de Jesus Zerbini (INCOR). Hospital das Clínicas da Faculdade de Medicina da USP (HCFMUSP). Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil |
| Local de pesquisa : | Aix-Marseille Université (AMU), France |
Abstract The endoplasmic reticulum chaperone Protein disulfide Isomerase A1 (PDI) emerges as a novel organizer of redox signaling processes involving Nox NADPH Oxidases. Recent evidences from our group suggest that mechanisms for these effects involve direct PDI convergence with RhoGTPases such as Rac1 and their regulator RhoGDI. In this context, PDI and related proteins are regulators of cytoskeletal dynamics. Using proteomic tools, we identified that another protein potentially involved in these interactions is the actin-binding nucleator profilin-1(PFN1). The experiments necessary to advance into understanding the in vivo functional relevance of PDI/PFN1 interactions (plus possible additional interactions with actin and Rac1) will require tools that are either not available or not entirely dominated by our group. Such techniques include high-resolution confocal microscopy, FRET and FLIM. Moreover, additional in vitro assays of actin polymerization and some experiments related to confirmation of protein complexation through biophysical techniques are proposed. The main objective of our BEPE scholarship is to validate and to address the pattern of PDI/PFN1 interactions in physiologically relevant model(s) involving cytoskeletal dynamics in endothelial cell migration. The specific aims are: 1) To investigate the functional implications of PDI and PFN1, as well as specific mutations targeted to disrupt their interaction, in an endothelial cell migration model through gain and loss of function experiments; 2)To analyze direct PDI/PFN1 protein interaction in live cells using microscopy tools; 3)To study PDI/PFN1 interaction during in vitro actin polymerization; 4) To determine the in vitro biochemical and physical parameters of relevant protein-protein interactions by microcalorimetry and analytical ultracentrifugation. Such experiments will contribute to advance into characterizing at a molecular level a model of PDI-controlled cytoskeletal dynamics, with considerable physiological implications. (AU) | |