The PTMs crosstalk in lung epithelial cells during polarization and infection

• Grant number: 23/08586-6
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
• Start date: May 15, 2024
• End date: September 23, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Giuseppe Palmisano
• Grantee: Deivid Martins Santos
• Supervisor: Martin Rossel Larsen
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: University of Southern Denmark (SDU), Denmark
• Associated to the scholarship: 21/14179-9 - The role of the endoplasmic reticulum in viral and parasitic virulence, BP.DD

Abstract

Post-translational modifications (PTMs) modulate proteins' structure, function, subcellular localization, and protein-protein interaction, playing a fundamental role in maintaining cellular homeostasis and are regulated during the pathogenesis of several diseases. N and O-linked glycan sialylation, phosphorylation and cysteine oxidation comprise mammalian cells' most studied PTMs. The recent development of analytical and computational methods has allowed identifying and quantifying thousands of modified proteins at specific sites with a fine-tuned stoichiometry. Enrichment techniques have allowed the isolation of low-abundant modified proteins/peptides for subsequent mass spectrometry-based analysis. In cell culture, novel models have been used like the Air-liquid interface (ALI) using the immortalized Callu-3 cell line. In this cell culture model is possible to mimic the normal lung epithelium and have been used in studies of lung infections e drug development. The main aim of this project is to identify and quantify PTMs at systems level and their regulation upon perturbation. This project will analyze the modulation and crosstalk of sialylated, phosphorylated and reversibly oxidized cysteine modifications during Calu-3 cell polarization and SARS-CoV-2 infection in an air-liquid interface (ALI) culture model. Previous data from our laboratory have revealed the regulation of kinases/phosphatases, glycosyltransferases, and redox enzymes during cellular polarization. Moreover, our study identified a different susceptibility to SARS-CoV-2 infection in the ALI model compared to the classic 2D cell model. Due to that, the main research questions are: 1) how PTMs (sialylation, phosphorylation and reversible modified cysteines) are regulated during polarization and viral infection? 2) are there specific protein hubs that could be targeted? Together with the expertise of Prof. Martin Larsen, this BEPE fellowship project will allow us to answer these questions and fill the knowledge gap in this field.